2,4-pyrimidinediamine compounds and uses as anti-proliferative agents

ABSTRACT

The present invention provides 2,4-pyrimidinediamine compounds having antiproliferative activity, compositions comprising the compounds and methods of using the compounds to inhibit cellular proliferation and to treat proliferative diseases such as tumorigenic cancers.

1. CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. application Ser. No.12/981,094, filed Dec. 29, 2010, which is a continuation of U.S.application Ser. No. 11/567,820, filed Dec. 7, 2006, which is acontinuation of U.S. patent application Ser. No. 10/913,270, filed Aug.6, 2004, now abandoned, Which claims benefit to U.S. ProvisionalApplication No. 60/494,008, filed Aug. 7, 2003 and U.S. ProvisionalApplication No. 60/572,534, filed May 18, 2004. Each of the foregoingapplications is incorporated by reference in its entirety.

2. FIELD

The present invention relates to 2,4 pyrimidinediamine compounds thatexhibit antiproliferative activity, prodrugs of the compounds,intermediates and methods of synthesis for making the compounds and/orprodrugs, pharmaceutical compositions comprising the compounds and theuse of the compounds in a variety of contexts, including for thetreatment of proliferative disorders, such as, for example, tumors andcancers.

3. BACKGROUND

Cancer is a group of varied diseases characterized by uncontrolledgrowth and spread of abnormal cells. Generally, all types of cancersinvolve some abnormality in the control of cell growth and division. Thepathways regulating cell division and/or cellular communication becomealtered in cancer cells such that the effects of these regulatorymechanisms in controlling and limiting cell growth fails or is bypassed.Through successive rounds of mutation and natural selection, a group ofabnormal cells, generally originating from a single mutant cell,accumulates additional mutations that provide selective growth advantageover other cells, and thus evolves into a cell type that predominates inthe cell mass. This process of mutation and natural selection isenhanced by genetic instability displayed by many types of cancer cells,an instability which is gained either from somatic mutations or byinheritance from the germ line. The enhanced mutability of cancerouscells increases the probability of their progression towards formationof malignant cells. As the cancer cells further evolve, some becomelocally invasive and then metastasize to colonize tissues other than thecancer cell's tissue of origin. This property along with theheterogeneity of the tumor cell population makes cancer a particularlydifficult disease to treat and eradicate.

Traditional cancer treatments take advantage of the higher proliferativecapacity of cancer cells and their increased sensitivity to DNA damage.Ionizing radiation, including γ-rays and x-rays, and cytotoxic agents,such as bleomycin, cis-platin, vinblastine, cyclophosphamide,5′-fluorouracil, and methotrexate rely upon a generalized damage to DNAand destabilization of chromosomal structure which eventually lead todestruction of cancer cells. These treatments are particularly effectivefor those types of cancers that have defects in cell cycle checkpoint,which limits the ability of these cells to repair damaged DNA beforeundergoing cell division. The non-selective nature of these treatments,however, often results in severe and debilitating side effects. Thesystemic use of these drugs may result in damage to normally healthyorgans and tissues, and compromise the long term health of the patient.

Although more selective chemotherapeutic treatments have been developedbased on knowledge of how cancer cells develop, for example, theanti-estrogen compound tamoxifen, the effectiveness of allchemotherapeutic treatments are subject to development of resistance tothe drugs. In particular, the increased expression of cell membranebound transporters, such as MdrI, produces a multidrug resistancephenotype characterized by increased efflux of drugs from the cell.These types of adaptation by cancer cells severely limit theeffectiveness of certain classes of chemotherapeutic agents.Consequently, identification of other chemotherapeutic agents iscritical for establishing therapies effective for attacking theheterogeneous nature of proliferative disease and for overcoming anyresistance that may develop over the course of therapy with othercompounds. Moreover, use of combinations of chemotherapeutic agents withdiffering properties and cellular targets increases the effectiveness ofchemotherapy and limits the generation of drug resistance.

4. SUMMARY

In one aspect, the present invention provides 2,4-pyrimidinediaminecompounds that exhibit antiproliferative activity against a variety ofdifferent cell types, including a variety of different types of tumorcells. The compounds are generally 2,4-pyrimidinediamine compoundsaccording to structural formula (I):

including salts, hydrates, solvates and N-oxides thereof, wherein:

-   -   L¹ and L² are each, independently of one another, selected from        a lower alkyldiyl linker, a lower alkylene linker and a covalent        bond;    -   R² is selected from the group consisting of lower alkyl        optionally substituted with an R^(b) group.

where Y is NH, O or CH₂;

-   -   R^(2′) is hydrogen, methyl or lower alkyl;    -   R^(4′) is hydrogen, methyl or lower alkyl;    -   R⁴ is selected from the group consisting of lower alkyl        optionally monosubstituted with an R^(a) or R^(b) group, lower        cycloalkyl optionally monosubstituted with an R^(a) or R^(b)        group, lower cycloheteroalkyl optionally substituted at one or        more ring carbon and/or heteroatoms with an R^(a) or R^(b)        group, —(CR^(a)R^(a))_(n)—R^(b),

where D is —(CR⁷R⁷)_(m)—,

where Z¹ is N or CH and Z² is O, S, NH, S(O) or S(O)₂;

-   -   R⁵ is selected from the group consisting of halo, fluoro and        —CF₃;    -   R⁶ is hydrogen;    -   each R⁷ is independently selected from the group consisting of        hydrogen, methyl, lower alkyl and halo;    -   each R⁸ is independently selected from the group consisting of        hydrogen, lower alkyl, —(CH₂)_(n)—OH, —OR^(a),        —(CH₂)_(n)—NR^(c)R^(c), —O(CH₂)_(n)—R^(a), —O(CH₂)_(n)—R^(b),        —C(O)OR^(a), —C(S)OR^(a,) , halo, —CF₃ and —OCF₃;    -   each R⁹ is independently selected from the group consisting of        hydrogen, lower alkyl, —OR^(a), —(CH₂)_(n)—NR^(c)R^(c),        —O(CH₂)_(n)—R^(a), —O(CH₂)_(n)—R^(b), —C(O)—NR^(c)R^(c),        —C(S)—NR^(c)R^(c), —S(O)₂—NR^(c)R^(c), —NHC(O)R^(a),        —NHC(S)R^(a), —C(O)—NH—(CH₂)_(n)—NR^(c)R^(c),        —C(S)—NH—(CH₂)_(n)—NR^(c)R^(c), halo, —CF₃, —OCF₃,

-   -   each R¹⁰ is independently selected from the group consisting of        hydrogen, lower alkyl, —(CH₂)_(n)—OH, —(CH₂)_(n)—NR^(c)R^(c),        —OR^(a), —O(CH₂)_(n)—R^(a), —O(CH₂)_(n)—R^(b), halo, —CF₃,        —OCF₃,

-   -   each R¹¹ is independently selected from the group consisting of        —OR^(a), —NR^(c)R^(c) and —NR^(a)R^(d);    -   each R¹² is independently selected from the group consisting of        lower alkyl, arylalkyl, —OR^(a), —NR^(c)R^(c), —C(O)R^(a),        —C(O)OR^(a) and —C(O)NR^(c)R^(c);    -   each R¹³ is independently selected from the group consisting of        lower alkyl, hydroxy, lower alkoxy, methoxy, —C(O)NR^(c)R^(c)        and —C(O)NH₂;    -   each R¹⁵ is independently selected from the group consisting of        hydrogen, lower alkyl, lower cycloakyl and phenyl;    -   each R¹⁶ is independently selected from the group consisting of        hydrogen, methyl, lower alkyl, lower cycloalkyl, lower branched        alkyl and lower cycloalkylmethyl;    -   each R¹⁷ is independently selected from the group consisting of        hydrogen, lower alkyl, methyl and R^(d) or, alternatively, R¹⁷        may be taken together with R¹⁸ to form an oxo (═O) group;    -   each R¹⁸ is independently selected from the group consisting of        hydrogen, lower alkyl and methyl or, alternatively, R¹⁸ may be        taken together with R¹⁷ to form an oxo (═O) group;    -   each R¹⁹ is independently selected form the group consisting of        hydrogen, lower alkyl, methyl and R^(d);    -   each R²⁰ is independently selected from the group consisting of        hydrogen, lower alkyl, methyl and R^(d);    -   each m is independently an integer from 1 to 3;    -   each n is independently an integer from 1 to 3;    -   each R^(a) is independently selected from the group consisting        of hydrogen, lower alkyl, lower cycloalkyl, lower        cycloalkylalkyl, phenyl and benzyl;    -   each R^(b) is independently selected from the group consisting        of —OR^(a), —CF₃, —OCF₃, —NR^(c)R^(c), —C(O)R^(a), —C(S)R^(a),        —C(O)OR^(a), —C(S)OR^(a), —C(O)NR^(c)R^(c), —C(S)NR^(c)R^(c),        —S(O)₂NR^(c)R^(c), —C(O)NR^(a)R^(d), —C(S)NR^(a)R^(d) and        —S(O)₂NR^(a)R^(d);    -   each R^(c) is independently selected from the group consisting        of hydrogen, lower alkyl and lower cycloalkyl, or,        alternatively, two R^(c)s may be taken together with the        nitrogen atom to which they are bonded to form a 5-7 membered        saturated ring which optionally includes 1-2 additional        heteroatomic groups selected from O, NR^(a), NR^(a)—C(O)R^(a),        NR^(a)—C(O)OR^(a) and NR^(a)—C(O)NR^(a); and    -   each R^(d) is independently selected from lower        mono-hydroxyalkyl and lower di-hydroxyalkyl.

In another aspect, the present invention provides prodrugs of the2,4-pyrimidinediamine compounds. Such prodrugs may be active in theirprodrug form, or may be inactive until converted under physiological orother conditions of use to an active drug form. In the prodrugs, one ormore functional groups of the 2,4-pyrimidinediamine compounds areincluded in promoieties that cleave from the molecule under theconditions of use, typically by way of hydrolysis, enzymatic cleavage orsome other cleavage mechanism, to yield the functional groups. Forexample, primary or secondary amino groups may be included in an amidepromoiety that cleaves under conditions of use to generate the primaryor secondary amino group. Thus, the prodrugs include special types ofprotecting groups, termed “progroups,” masking one or more functionalgroups of the 2,4-pyrimidinediamine compounds that cleave under theconditions of use to yield an active 2,4-pyrimidinediamine drugcompound. Functional groups within the 2,4-pyrimidinediamine compoundsthat may be masked with progroups for inclusion in a promoiety include,but are not limited to, amines (primary and secondary), hydroxyls,sulfanyls (thiols), carboxyls, carbonyls, phenols, catechols, diols,alkynes, phosphates, etc. Myriad progroups suitable for masking suchfunctional groups to yield promoieties that are cleavable under thedesired conditions of use are known in the art. All of these progroups,alone or in combinations, may be included in the prodrugs. Specificexamples of promoieties that yield primary or secondary amine groupsthat can be included in the prodrugs include, but are not limited toamides, carbamates, imines, ureas, phosphenyls, phosphoryls andsulfenyls. Specific examples of promoieties that yield sulfanyl groupsthat can be included in the prodrugs include, but are not limited to,thioethers, for example S-methyl derivatives (monothio, dithio, oxythio,aminothio acetals), silyl thioethers, thioesters, thiocarbonates,thiocarbamates, asymmetrical disulfides, etc. Specific examples ofpromoieties that cleave to yield hydroxyl groups that can be included inthe prodrugs include, but are not limited to, sulfonates, esters andcarbonates. Specific examples of promoieties that yield carboxyl groupsthat can be included in the prodrugs include, but are not limited to,esters (including silyl esters, oxamic acid esters and thioesters),amides and hydrazides.

In another aspect, the present invention provides compositionscomprising one or more 2,4-pyrimidinediamine compounds and/or prodrugsand an appropriate carrier, excipient and/or diluent. The exact natureof the carrier, excipient and/or diluent will depend upon the desireduse for the composition, and may range from being suitable or acceptablefor veterinary uses to being suitable or acceptable for human use.

The 2,4-pyrimidinediamine compounds are potent inhibitors ofproliferation abnormal cells, such as tumor cell proliferation, in invitro assays. Thus, in still another aspect, the present inventionprovides methods of inhibiting proliferation of abnormal cells, inparticular tumor cells. The method generally involves contacting anabnormal cell such as a tumor cells with an amount of a2,4-pyrimidinediamine compound or prodrug, or an acceptable salt,hydrate, solvate, N-oxide and/or composition thereof, effective toinhibit its proliferation. The method may be practiced in in vitrocontexts or in in vivo contexts as a therapeutic approach towards thetreatment or prevention of proliferative disorders, such as tumorigeniccancers.

In still another aspect, the present invention provides methods oftreating proliferative disorders. The methods may be practiced inanimals in veterinary contexts or in humans. The methods generallyinvolve administering to an animal or human subject an amount of a2,4-pyrimidinediamine compound or prodrug, or an acceptable salt,hydrate, solvate, N-oxide and/or composition thereof, effective to treatthe disorder. Proliferative disorders that can be treated according tothe methods include, but are not limited to, tumorigenic cancers.

Other aspects of the present invention include, but are not limited to,intermediates and methods useful for synthesizing the compound andprodrugs, as will be described in more detail herein below.

5. DETAILED DESCRIPTION 5.1 Definitions

As used herein, the following terms are intended to have the followingmeanings.

“Alkyl” by itself or as part of another substituent refers to asaturated or unsaturated branched, straight-chain or cyclic monovalenthydrocarbon radical having the stated number of carbon atoms (i.e.,C1-C6 means one to six carbon atoms) that is derived by the removal ofone hydrogen atom from a single carbon atom of a parent alkane, alkeneor alkyne. Typical alkyl groups include, but are not limited to, methyl;ethyls such as ethanyl, ethenyl, ethynyl; propyls such as propan-1-yl,propan-2-yl, cyclopropan-1-yl, prop-1-en-1-yl, prop-1-en-2-yl,prop-2-en-1-yl, cycloprop-1-en-1-yl; cycloprop-2-en-1-yl,prop-1-yn-1-yl, prop-2-yn-1-yl, etc.; butyls such as butan-1-yl,butan-2-yl, 2-methyl-propan-1-yl, 2-methyl-propan-2-yl, cyclobutan-1-yl,but-1-en-1-yl, but-1-en-2-yl, 2-methyl-prop-1-en-1-yl, but-2-en-1-yl,but-2-en-2-yl, buta-1,3-dien-1-yl, buta-1,3-dien-2-yl,cyclobut-1-en-1-yl, cyclobut-1-en-3-yl, cyclobuta-1,3-dien-1-yl,but-1-yn-1-yl, but-1-yn-3-yl, but-3-yn-1-yl, etc.; and the like. Wherespecific levels of saturation are intended, the nomenclature “alkanyl,”“alkenyl” and/or “alkynyl” is used, as defined below. “Lower alkyl”refers to alkyl groups having from 1 to 8 carbon atoms.

“Alkanyl” by itself or as part of another substituent refers to asaturated branched, straight-chain or cyclic alkyl derived by theremoval of one hydrogen atom from a single carbon atom of a parentalkane. Typical alkanyl groups include, but are not limited to,methanyl; ethanyl; propanyls such as propan-1-yl, propan-2-yl(isopropyl), cyclopropan-1-yl, etc.; butanyls such as butan-1-yl,butan-2-yl (sec-butyl), 2-methyl-propan-1-yl (isobutyl),2-methyl-propan-2-yl (t-butyl), cyclobutan-1-yl, etc.; and the like.

“Alkenyl” by itself or as part of another substituent refers to anunsaturated branched, straight-chain or cyclic alkyl having at least onecarbon-carbon double bond derived by the removal of one hydrogen atomfrom a single carbon atom of a parent alkene. The group may be in eitherthe cis or trans conformation about the double bond(s). Typical alkenylgroups include, but are not limited to, ethenyl; propenyls such asprop-1-en-1-yl, prop-1-en-2-yl, prop-2-en-1-yl, prop-2-en-2-yl,cycloprop-1-en-1-yl; cycloprop-2-en-1-yl; butenyls such asbut-1-en-1-yl, but-1-en-2-yl, 2-methyl-prop-1-en-1-yl, but-2-en-1-yl,but-2-en-2-yl, buta-1,3-dien-1-yl, buta-1,3-dien-2-yl,cyclobut-1-en-1-yl, cyclobut-1-en-3-yl, cyclobuta-1,3-dien-1-yl, etc.;and the like.

“Alkynyl” by itself or as part of another substituent refers to anunsaturated branched, straight-chain or cyclic alkyl having at least onecarbon-carbon triple bond derived by the removal of one hydrogen atomfrom a single carbon atom of a parent alkyne. Typical alkynyl groupsinclude, but are not limited to, ethynyl; propynyls such asprop-1-yn-1-yl, prop-2-yn-1-yl, etc.; butynyls such as but-1-yn-1-yl,but-1-yn-3-yl, but-3-yn-1-yl, etc.; and the like.

“Alkyldiyl” by itself or as part of another substituent refers to asaturated or unsaturated, branched, straight-chain or cyclic divalenthydrocarbon group having the stated number of carbon atoms (i.e., C1-C6means from one to six carbon atoms) derived by the removal of onehydrogen atom from each of two different carbon atoms of a parentalkane, alkene or alkyne, or by the removal of two hydrogen atoms from asingle carbon atom of a parent alkane, alkene or alkyne. The twomonovalent radical centers or each valency of the divalent radicalcenter can form bonds with the same or different atoms. Typicalalkyldiyl groups include, but are not limited to, methandiyl; ethyldiylssuch as ethan-1,1-diyl, ethan-1,2-diyl, ethen-1,1-diyl, ethen-1,2-diyl;propyldiyls such as propan-1,1-diyl, propan-1,2-diyl, propan-2,2-diyl,propan-1,3-diyl, cyclopropan-1,1-diyl, cyclopropan-1,2-diyl,prop-1-en-1,1-diyl, prop-1-en-1,2-diyl, prop-2-en-1,2-diyl,prop-1-en-1,3-diyl, cycloprop-1-en-1,2-diyl, cycloprop-2-en-1,2-diyl,cycloprop-2-en-1,1-diyl, prop-1-yn-1,3-diyl, etc.; butyldiyls such as,butan-1,1-diyl, butan-1,2-diyl, butan-1,3-diyl, butan-1,4-diyl,butan-2,2-diyl, 2-methyl-propan-1,1-diyl, 2-methyl-propan-1,2-diyl,cyclobutan-1,1-diyl; cyclobutan-1,2-diyl, cyclobutan-1,3-diyl,but-1-en-1,1-diyl, but-1-en-1,2-diyl, but-1-en-1,3-diyl,but-1-en-1,4-diyl, 2-methyl-prop-1-en-1,1-diyl,2-methanylidene-propan-1,1-diyl, buta-1,3-dien-1,1-diyl,buta-1,3-dien-1,2-diyl, buta-1,3-dien-1,3-diyl, buta-1,3-dien-1,4-diyl,cyclobut-1-en-1,2-diyl, cyclobut-1-en-1,3-diyl, cyclobut-2-en-1,2-diyl,cyclobuta-1,3-dien-1,2-diyl, cyclobuta-1,3-dien-1,3-diyl,but-1-yn-1,3-diyl, but-1-yn-1,4-diyl, buta-1,3-diyn-1,4-diyl, etc.; andthe like. Where specific levels of saturation are intended, thenomenclature alkanyldiyl, alkenyldiyl and/or alkynyldiyl is used. Whereit is specifically intended that the two valencies are on the samecarbon atom, the nomenclature “alkylidene” is used. A “lower alkyldiyl”is an alkyldiyl group having from 1 to 6 carbon atoms. In preferredembodiments the alkyldiyl groups are saturated acyclic alkanyldiylgroups in which the radical centers are at the terminal carbons, e.g.,methandiyl (methano); ethan-1,2-diyl (ethano); propan-1,3-diyl(propano); butan-1,4-diyl (butano); and the like (also referred to asalkylenes, defined infra).

“Alkylene” by itself or as part of another substituent refers to astraight-chain saturated or unsaturated alkyldiyl group having twoterminal monovalent radical centers derived by the removal of onehydrogen atom from each of the two terminal carbon atoms ofstraight-chain parent alkane, alkene or alkyne. The locant of a doublebond or triple bond, if present, in a particular alkylene is indicatedin square brackets. Typical alkylene groups include, but are not limitedto, methylene (methano); ethylenes such as ethano, etheno, ethyno;propylenes such as propano, prop[1]eno, propa[1,2]dieno, prop[1]yno,etc.; butylenes such as butano, but[1]eno, but[2]eno, buta[1,3]dieno,but[1]yno, but[2]yno, buta[1,3]diyno, etc.; and the like. Where specificlevels of saturation are intended, the nomenclature alkano, alkenoand/or alkyno is used. In preferred embodiments, the alkylene group is(C1-C6) or (C1-C3) alkylene. Also preferred are straight-chain saturatedalkano groups, e.g., methano, ethano, propano, butano, and the like.

“Cycloalkyl” by itself or as part of another substituent refers to acyclic version of an “alkyl” group. Typical cycloalkyl groups include,but are not limited to, cyclopropyl; cyclobutyls such as cyclobutanyland cyclobutenyl; cyclopentyls such as cyclopentanyl and cyclopentenyl;cyclohexyls such as cyclohexanyl and cyclohexenyl; and the like. “Lowercycloalkyl” refers to a cycloalkyl group having from 3 to 8 ring carbonatoms.

“Cycloalkylalkyl” by itself or as part of another substitutent refers toan alkyl group that comprises a linear or branched portion and a cyclicportion. Typical cycloalkylalkyl groups include, but are not limited to,cyclopropylmethyl, 1-cycopropyleth-1-yl, 2-cyclopropyleth-1-yl,cyclobutylmethyl, 1-cycobytyleth-1-yl, 2-cyclobutyleth-1-yl,cyclopentylmethyl, 1-cycopentyleth-1-yl, 2-cyclopentyleth-1-yl,cyclohexylmethyl, 1-cycohexyleth-1-yl, 2-cyclohexleth-1-yl, and thelike. “Lower cycloalkylalkyl” refers to a cycloalkylalkyl group in whichthe linear or branched portion contains from 1 to 4 carbon atoms and thecyclic portion contains from 3 to 8 carbon atoms.

“Heteroalkyl” by itself or as part of another substituent refers to analkyl group in which at least one of the carbon atoms is replaced with aheteroatom, for example, a heteroatom selected from O, S and N. Inheteroalkyl groups including more than one heteroatom, the heteroatomsmay be the same or they may be different. Like an alkyl group, aheteroalkyl can be linear, branched or cyclic in structure, and can besaturated or unsaturated. Typical heteroalkyl groups include, but arenot limited to, —CH₂—O—CH₂—, —CH₂—S—CH₂—, —CH₂—NH—CH₂—,—CH₂—N(CH₃)—CH₂—,

—CH₂CH₂—O—CH₂—, —CH₂CH₂—S—CH₂—, —CH₂CH₂—NH—CH₂—, —CH₂CH₂—N(CH₃)—CH₂—,

—CH₂CH(CH₃)—O—CH₂—, —CH₂CH(CH₃)—S—CH₂, —CH₂CH(CH₁)—NH—CH₂—,—CH₂CH(CH₃)—N(CH₃)—CH₂—,

—CH═CH—O—CH₂—, —CH═CH—S—CH₂—, —CH═CH—NH—CH₂—, —CH═CH—N(CH₃)—CH₂—,

—C≡C—O—CH₂—, —C≡C—S—CH₂—, —C≡C—NH—CH₂—, —C≡C—N(CH₃)—CH₂—, and the like.Where specific levels of saturation are intended, the nomenclature“heteroalkanyl,” “heteroalkenyl,” and heteroalkynyl” is used. “Lowerheteroalkyl” refers to a heteroalkyl group having from 1 to 8 carbon andheteroatoms.

“Cycloheteroalkyl” by itself or as part of another substituent refers toa cyclic version of a heteroalkyl. Typical examples of cycloheteroalkylgroups include, but are not limited to,

and the like. “Lower cycloheteroalkyl” refers to a cycloheteroalkylgroup having from 3 to 8 ring atoms.

“Parent Aromatic Ring System” refers to an unsaturated cyclic orpolycyclic ring system having a conjugated π electron system.Specifically included within the definition of “parent aromatic ringsystem” are fused ring systems in which one or more of the rings arearomatic and one or more of the rings are saturated or unsaturated, suchas, for example, fluorene, indane, indene, phenalene,tetrahydronaphthalene, etc. Typical parent aromatic ring systemsinclude, but are not limited to, aceanthrylene, acenaphthylene,acephenanthrylene, anthracene, azulene, benzene, chrysene, coronene,fluoranthene, fluorene, hexacene, hexaphene, hexalene, indacene,s-indacene, indane, indene, naphthalene, octacene, octaphene, octalene,ovalene, penta-2,4-diene, pentacene, pentalene, pentaphene, perylene,phenalene, phenanthrene, picene, pleiadene, pyrene, pyranthrene,rubicene, tetrahydronaphthalene, triphenylene, trinaphthalene, and thelike, as well as the various hydro isomers thereof.

“Aryl” by itself or as part of another substituent refers to amonovalent aromatic hydrocarbon group having the stated number of carbonatoms (i.e., C5-C15 means from 5 to 15 carbon atoms) derived by theremoval of one hydrogen atom from a single carbon atom of a parentaromatic ring system. Typical aryl groups include, but are not limitedto, groups derived from aceanthrylene, acenaphthylene,acephenanthrylene, anthracene, azulene, benzene, chrysene, coronene,fluoranthene, fluorene, hexacene, hexaphene, hexalene, as-indacene,s-indacene, indane, indene, naphthalene, octacene, octaphene, octalene,ovalene, penta-2,4-diene, pentacene, pentalene, pentaphene, perylene,phenalene, phenanthrene, picene, pleiadene, pyrene, pyranthrene,rubicene, triphenylene, trinaphthalene, and the like, as well as thevarious hydro isomers thereof. In preferred embodiments, the aryl groupis (C5-C15) aryl, with (C5-C10) being even more preferred. Particularlypreferred aryls are phenyl and naphthyl.

“Halogen” or “Halo” by themselves or as part of another substituent,unless otherwise stated, refer to fluoro, chloro, bromo and iodo.

“Haloalkyl” by itself or as part of another substituent refers to analkyl group in which one or more of the hydrogen atoms is replaced witha halogen. Thus, the term “haloalkyl” is meant to includemonohaloalkyls, dihaloalkyls, trihaloalkyls, etc. up to perhaloalkyls.For example, the expression “(C1-C2) haloalkyl” includes fluoromethyl,difluoromethyl, trifluoromethyl, 1-fluoroethyl, 1,1-difluoroethyl,1,2-difluoroethyl, 1,1,1-trifluoroethyl, perfluoroethyl, etc.

The above-defined groups may include prefixes and/or suffixes that arecommonly used in the art to create additional well-recognizedsubstituent groups. As non-limiting examples, “alkyloxy” or “alkoxy”refers to a group of the formula —OR, “alkylamine” refers to a group ofthe formula —NHR and “dialkylamine” refers to a group of the formula—NRR, where each R is independently an alkyl. As another non-limitingexample, “haloalkoxy” or “haloalkyloxy” refers to a group of the formula—OR′, where R′ is a haloalkyl.

“Prodrug” refers to a derivative of an active 2,4-pyrimidinediaminecompound (drug) that may require a transformation under the conditionsof use, such as within the body, to release the active2,4-pyrimidinediamine drug. Prodrugs are frequently, but notnecessarily, pharmacologically inactive until converted into the activedrug. Prodrugs are typically obtained by masking a functional group inthe 2,4-pyrimidinediamine drug believed to be in part required foractivity with a progroup (defined below) to form a promoiety whichundergoes a transformation, such as cleavage, under the specifiedconditions of use to release the functional group, and hence the active2,4-pyrimidinediamine drug. The cleavage of the promoiety may proceedspontaneously, such as by way of a hydrolysis reaction, or it may becatalyzed or induced by another agent, such as by an enzyme, by light,by acid or base, or by a change of or exposure to a physical orenvironmental parameter, such as a change of temperature. The agent maybe endogenous to the conditions of use, such as an enzyme present in thecells to which the prodrug is administered or the acidic conditions ofthe stomach, or it may be supplied exogenously.

A wide variety of progroups, as well as the resultant promoieties,suitable for masking functional groups in the active2,4-pyrimidinediamines compounds to yield prodrugs are well-known in theart. For example, a hydroxyl functional group may be masked as asulfonate, ester or carbonate promoiety, which may be hydrolyzed in vivoto provide the hydroxyl group. An amino functional group may be maskedas an amide, carbamate, imine, urea, phosphenyl, phosphoryl or sulfenylpromoiety, which may be hydrolyzed in vivo to provide the amino group. Acarboxyl group may be masked as an ester (including silyl esters andthioesters), amide or hydrazide promoiety, which may be hydrolyzed invivo to provide the carboxyl group. Other specific examples of suitableprogroups and their respective promoieties will be apparent to those ofskill in the art.

“Progroup” refers to a type of protecting group that, when used to maska functional group within an active 2,4-pyrimidinediamine drug to form apromoiety, converts the drug into a prodrug. Progroups are typicallyattached to the functional group of the drug via bonds that arecleavable under specified conditions of use. Thus, a progroup is thatportion of a promoiety that cleaves to release the functional groupunder the specified conditions of use. As a specific example, an amidepromoiety of the formula —NH—C(O)CH₃ comprises the progroup —C(O)CH₃.

“Proliferative disorder” refers to a disease or disorder characterizedby aberrant cell proliferation, for example where cells divide more thantheir counterpart normal cells. The aberrant proliferation may be causedby any mechanism of action or combination of mechanisms of action. Forexample, the cell cycle of one or more cells may be affected such thatcell(s) divide more frequently than their counterpart normal cells, oralternatively, one or more cells may bypass inhibitory signals whichwould normally limit their number of divisions. Proliferative diseasesinclude, but are not limited to, slow or fast growing tumors andcancers.

“Antiproliferative compound” refers to a compound that inhibits theproliferation of a cell as compared to an untreated control cell of asimilar type. The inhibition can be brought about by any mechanism orcombination of mechanisms, and may operate to inhibit proliferationcytostatically or cytotoxically. As a specific example, inhibition asused herein includes, but is not limited to, arrest of cell division, areduction in the rate of cell division, proliferation and/or growthand/or induction of cell death.

“Pharmaceutically effective amount” or “therapeutically effectiveamount” refers to an amount of a compound sufficient to treat aspecified disorder or disease or one or more of its symptoms and/or toprevent the occurrence of the disease or disorder. In reference totumorigenic proliferative disorders, a pharmaceutically ortherapeutically effective amount comprises an amount sufficient to,among other things, cause the tumor to shrink or to decrease the growthrate of the tumor.

5.2 Antiproliferative 2,4-Pyrimidinediamine Compounds

The antiproliferative compounds are generally 2,4-pyrimidinediaminecompounds according to structural formula (I):

including salts, hydrates, solvates and N-oxides thereof, wherein:

-   -   L¹ and L² are each, independently of one another, selected from        a lower alkyldiyl linker, a lower alkylene linker and a covalent        bond;    -   R² is selected from the group consisting of lower alkyl        optionally substituted with an R^(b) group,

where Y is NH, 0 or CH₂;

-   -   R^(2′) is hydrogen, methyl or lower alkyl;    -   R^(4′) is hydrogen, methyl or lower alkyl;    -   R⁴ is selected from the group consisting of lower alkyl        optionally monosubstituted with an R^(a) or R^(b) group, lower        cycloalkyl optionally monosubstituted with an R^(a) or R^(b)        group, lower cycloheteroalkyl optionally substituted at one or        more ring carbon and/or heteroatoms with an R^(a) or R^(b)        group, —(CR^(a)R^(a))_(n)—R^(b),

where D is —(CR⁷R⁷)_(m)—,

where Z¹ is N or CH and Z² is O, S, NH, S(O) or S(O)₂;

-   -   R⁵ is selected from the group consisting of halo, fluoro and        —CF₃;    -   R⁶ is hydrogen;    -   each R⁷ is independently selected from the group consisting of        hydrogen, methyl, lower alkyl and halo;    -   each R⁸ is independently selected from the group consisting of        hydrogen, lower alkyl, —(CH₂)_(n)—OH, —OR^(a),        —(CH₂)_(n)—NR^(c)R^(c), —O(CH₂)_(n)—R^(a), —O(CH₂)_(n)—R^(b),        —C(O)OR^(a), —C(S)OR^(a′) halo, —CF₃ and —OCF₃;    -   each R⁹ is independently selected from the group consisting of        hydrogen, lower alkyl, —OR^(a), —(CH₂)_(n)—NR^(c)R^(c),        —O(CH₂)_(n)—R^(a), —O(CH₂)_(n)—R^(b), —C(O)—NR^(c)R^(c),        —C(S)—NR^(c)R^(c), —S(O)₂—NR^(c)R^(c), —NHC(O)R^(a),        —NHC(S)R^(a), —C(O)—NH—(CH₂)_(n)—NR^(c)R^(c),        —C(S)—NH—(CH₂)_(n)—NR^(c)R^(c), halo, —CF₃, —OCF₃,

-   -   each R¹⁰ is independently selected from the group consisting of        hydrogen, lower alkyl, —(CH₂)_(n)—OH, —(CH₂)_(n)—NR^(c)R^(c),        —OR^(a), —O(CH₂)_(n)—R^(a), —O(CH₂)_(n)—R^(b), halo, —CF₃,        —OCF₃,

-   -   each R¹¹ is independently selected from the group consisting of        —OR^(a), —NR^(c)R^(c) and —NR^(a)R^(d);    -   each R¹² is independently selected from the group consisting of        lower alkyl, arylalkyl, —OR^(a), —NR^(c)R^(c), —C(O)R^(a),        —C(O)OR^(a) and —C(O)NR^(c)R^(c);    -   each R¹³ is independently selected from the group consisting of        lower alkyl, hydroxy, lower alkoxy, methoxy, —C(O)NR^(c)R^(c)        and —C(O)NH₂;    -   each R¹⁵ is independently selected from the group consisting of        hydrogen, lower alkyl, lower cycloakyl and phenyl;    -   each R¹⁶ is independently selected from the group consisting of        hydrogen, methyl, lower alkyl, lower cycloalkyl, lower branched        alkyl and lower cycloalkylmethyl;    -   each R¹⁷ is independently selected from the group consisting of        hydrogen, lower alkyl, methyl and R^(d) or, alternatively, R¹⁷        may be taken together with R¹⁸ to form an oxo (═O) group;    -   each R¹⁸ is independently selected from the group consisting of        hydrogen, lower alkyl and methyl or, alternatively, R¹⁸ may be        taken together with R¹⁷ to form an oxo (═O) group;    -   each R¹⁹ is independently selected form the group consisting of        hydrogen, lower alkyl, methyl and R^(d);    -   each R²⁰ is independently selected from the group consisting of        hydrogen, lower alkyl, methyl and R^(d);    -   each m is independently an integer from 1 to 3;    -   each n is independently an integer from 1 to 3;    -   each R^(a) is independently selected from the group consisting        of hydrogen, lower alkyl, lower cycloalkyl, lower        cycloalkylalkyl, phenyl and benzyl;    -   each R^(b) is independently selected from the group consisting        of —OR^(a), —CF₃, —OCF₃, —NR^(c)R^(c), —C(O)R^(a), —C(S)R^(a),        —C(O)OR^(a), —C(S)OR^(a), —C(O)NR^(c)R^(c), —C(S)NR^(c)R^(c),        —S(O)₂NR^(c)R^(c), —C(O)NR^(a)R^(d), —C(S)NR^(a)R^(d) and        —S(O)₂NR^(a)R^(d);    -   each R^(c) is independently selected from the group consisting        of hydrogen, lower alkyl and lower cycloalkyl, or,        alternatively, two R^(c)s may be taken together with the        nitrogen atom to which they are bonded to form a 5-7 membered        saturated ring which optionally includes 1-2 additional        heteroatomic groups selected from O, NR^(a), NR^(a)—C(O)R^(a),        NR^(a)—C(O)OR^(a) and NR^(a)—C(O)NR^(a); and    -   each R^(d) is independently selected from lower        mono-hydroxyalkyl and lower di-hydroxyalkyl.

An important class of compounds of structural formula (I) includescompounds in which L¹ and L² are each a covalent bond, such that thecompound is a 2,4-pyrimidine diamine according to structural formula(II):

including salts, hydrates, solvates and N-oxides thereof, wherein R₂,R^(2′), R⁴, R^(4′), R⁵ and R⁶ are as previously defined for structuralformula (I).

An important class of compounds of structural formulae (I) and/or (II)and the salts, hydrates, solvates and N-oxides thereof, includescompounds in which R⁵ is fluoro.

Another important class of compounds of structural formulae (I) and/or(II) and the salts, hydrates, solvates and N-oxides thereof, includescompounds in which R^(2′) is hydrogen.

Another important class of compounds of structural formulae (I) and/or(II) and the salts, hydrates, solvates and N-oxides thereof, includescompounds in which R^(2′) and R^(4′) are each, independently of oneanother, selected from hydrogen and methyl.

Another important class of compounds of structural formulae (I) and/or(II) and the salts, hydrates, solvates and N-oxides thereof, includescompounds in which R^(4′) is methyl.

Other important classes of compounds of structural formulae (I) and/or(II) include compounds according to structural formulae (III)-(V):

and salts, hydrates, solvates and N-oxides thereof, wherein R², R^(2′),R⁴ and R^(4′), are as previously defined for structural formula (I).

When R² and/or R⁴ is

in the compounds described herein, for example, the compounds ofstructural formulae (I)-(V), in some embodiments, R⁹ and R¹⁰ are notboth simultaneously lower alkoxy or methoxy. In other embodiments, R⁸,R⁹ and R¹⁰ are not each simultaneously lower alkoxy or methoxy. In stillother embodiments R⁸, R⁹ and R¹⁰ are each methoxy or lower alkoxy. Inyet other embodiments, R⁸ is selected from hydrogen, lower alkyl, loweralkoxy, —OR^(a), halo, —CF₃ and —OCF₃ and one of R⁹ or R¹⁰ is selectedfrom

In a specific embodiment, the other one of R⁹ or R¹⁰ is other than

In yet other embodiments, R⁸ is selected from hydrogen, lower alkyl,—OR^(a), halo, —CF₃ and —OCF₃ and one of R⁹ or R¹⁰ is selected from—OCH₂C(O)R^(a), —OCH₂C(O)OR^(a), —OCH₂C(O)NHR^(a), —OCH₂C(O)NHR^(d) and—OCH₂C(O)NR^(c)R^(c). In a specific embodiment, the other one of R⁹ orR¹⁰ is other than —OCH₂C(O)R^(a), OCH₂C(O)OR^(a), —OCH₂C(O)NHR^(a),—OCH₂C(O)NHR^(d) or —OCH₂C(O)NR^(c)R^(c).

In yet other embodiments, R⁸ and R⁹ are each, independently of oneanother, selected from hydrogen, lower alkyl, —OR^(a), halo, —CF₃ and—OR^(a) and R¹⁰ is

In yet other embodiments R⁹ is hydrogen and R⁸ and R¹⁰ are each,independently of one another, selected from the group consisting oflower alkyl, methyl, lower alkoxy, methoxy, —CF₃ and —OCF₃. Specificcombinations of R⁸ and R¹⁰ when R⁹ is hydrogen are as follows:

-   -   R⁸ and R¹⁰ are each the same lower alkyl or methyl;    -   R⁸ and R¹⁰ are each the same lower alkoxy or methoxy;    -   R⁸ is lower alkyl or methyl and R¹⁰ is —CF₃;    -   R⁸ is lower alkoxy or methoxy and R¹⁰ is —CF₃; and    -   R⁸ is lower alkyl or methyl and R¹⁰ is —OCF₃.

In still other embodiments, R⁹ is hydroxy, methoxy or chloro and R⁸ andR¹⁰ are each, independently of one another, selected from the groupconsisting of lower alkyl, methyl, lower alkoxy, methoxy and chloro.Specific combinations of R⁸, R⁹ and R¹⁰ according to this embodiment areas follows:

-   -   R⁸ and R¹⁰ are each methyl and R⁹ is hydroxy, methoxy or chloro;    -   R⁸ and R¹⁰ are each chloro and R⁹ is hydroxy or methoxy; and    -   R⁸ is chloro, R¹⁰ is methyl and R⁹ is hydroxy or methoxy.

When R² and R⁴ are each

in the compounds described herein, such as, for example, the compoundsof structural formulae (I)-(V), in some embodiments no more than one ofR⁸, R⁹ and R¹⁰ of the R⁴ phenyl is hydrogen unless at least one of R⁸,R⁹ or R¹⁰ of the R² phenyl is —O(CH₂)_(n)—NR^(c)R^(c),

In other embodiments, the substitution patterns of the R² and R⁴ phenylrings are different from each other such that the compound is not aN2,N4-bis(3,4,5-substituted phenyl)pyrimidinediamine.

In still other embodiments, the compound is a compound according tostructural formula (VI):

including the salts, hydrates, solvates and N-oxides thereof, whereinR^(4′), R⁸, R⁹ and R¹⁰ are as previously defined for structural formula(I).

In still other embodiments, the compound is a compound according tostructural formula (VII):

including the salts, hydrates, solvates and N-oxides thereof, whereinR¹⁶ and R²⁴ are each independently selected from hydrogen and methyl;R⁸ is selected from hydrogen, lower alkyl, methyl; hydroxy, loweralkoxy,methoxy, halo, chloro, trifluoromethyl and —CH₂OH;R⁹ is selected from hydrogen, lower alkyl, methyl, hydroxy, loweralkoxy, methoxy, halo, chloro, trifluoromethyl, trifluoromethoxy,—OCH₂C(O)NHR^(a),

R¹⁰ is selected from hydrogen, lower alkyl, methyl, hydroxy, loweralkoxy, methoxy, halo, chloro, trifluoromethyl, trifluoromethoxy,—OCH₂C(O)NHR^(a), —OCH₂C(O)OR^(a),

with the provisos that:(i) when R⁹ is

then R¹⁰ is other than

and(ii) when R¹⁰

then R⁹ is other than

and Z¹, R¹², R¹⁹, R²⁰, R^(a) are as defined formula (I),

In still other embodiments when R² and R⁴ are each

R⁸, R⁹ and R¹⁰ are selected such that each R² and R⁴ phenyl ring ismono-substituted.

In yet other embodiments when R² and R⁴ are each

R⁸, R⁹ and R¹⁰ are selected such that each R² and R⁴ phenyl ring isdi-substituted. In one specific embodiment, each R² and R⁴ phenyl ringis substituted with an ethylenedioxy acetal group.

In still other embodiments when R² and R⁴ are each

R¹⁰ of the R² or R⁴ ring is other than 1,3-oxazolyl or 1,3-oxazol-5-ylwhen the R⁸ and R⁹ of the same ring are each hydrogen. In one specificembodiment, R⁸, R⁹ and R¹⁰ are as defined in the preceeding sentencewhen R⁵ is fluoro and L² is a lower alkylene. In another specificembodiment, R² is other than 3-(1,3-oxazolyl)phenyl or3-(1,3-oxazol-5-yl)phenyl when R⁴ is 2-(trifluoromethyl)benzyl. Inanother specific embodiment, the compound is other thanN2-[3-(1,3-oxazolyl)phenyl]-N4-[2-trifluoromethy)lbenzyl]-5-fluoro-2,4-pyrimidinediamine.In compounds where R¹⁰ is an oxazolyl, the oxazolyl is not connected atthe 5 position. In a specific embodiment, the oxazole is connected atthe 2 position. In still another specific embodiment, the compound isnot any compound described in WO 03/040141, the disclosure of which isincorporated herein by reference.

When R² is

in the compounds described herein, such as, for example, the compoundsof structural formulae (I)-(VII), in some embodiments one of R⁹ or R¹⁰is selected from

where R¹² is as previously defined for structural formula (I) and theother one of R⁹ or R¹⁰ is other than

In still other embodiments, R⁸ is selected from hydrogen, lower alkyl,methyl, lower alkoxy, methoxy and halo and one of R⁹ or R¹⁰ is—OCH₂—R^(b), where R^(b) is selected from —C(O)NR^(a) and —C(O)NHR^(a),and the other one of R⁹ or R¹⁰ is selected from hydrogen, lower alkyl,methyl, lower alkoxy, methoxy and halo. In still other embodiments R² is

where R⁸ is hydrogen, fluoro or CF₃. In a specific embodiment, R² is

When R² is

in the compounds described herein, such as, for example, the compoundsaccording to structural formulae (I)-(V), in some embodiments, R⁸ and R⁹are each independently selected from hydrogen, lower alkyl, methyl,lower alkoxy, methoxy, halo and chloro. One specific embodiment, R⁸, R⁹and R¹³ are each independently selected from halo, lower alkyl, methyl,lower alkoxy, and methoxy. In another specific embodiment, R² isselected from

In some embodiments of compounds in which R² is

including any of the above-described specific embodiments, R⁴ isselected from

where D is as previously defined for structural formula (I).

When R² is 3,4,5-trimethoxyphenyl or 3,4,5-tri(loweralkoxy)phenyl in thecompounds described herein, such as, for example the compounds ofstructural formulae (I)-(V), in some embodiments, R⁴ is

where Z¹, Z² and R¹⁶, R¹⁷, R¹⁸, R¹⁹ and R²⁰ are as previously describedfor structural formula (I).

When R⁹ or R¹⁰ is

in the compounds described herein, such as, for example, the compoundsof structural formulae (I)-(VII), in some embodiments R¹² is methyl. Inother embodiments, R¹² is —C(O)R^(a) or —C(O)OR^(a), where R^(a) islower alkyl, ethyl or methyl.

When R² is

in the compounds described herein, such as, for example, the compoundsof structural formulae (I)-(V), in some embodiments (i) each R¹³ is,independently or the other, selected from lower alkyl, methyl, hydroxy,lower alkoxy and methoxy; and/or (ii) R⁴ is selected from

where Z¹, Z² and R¹⁶, R¹⁷, R¹⁸, R¹⁹ and R²⁰ are as previously describedfor structural formula (I). In a specific embodiment, R⁴ is

where Z¹ is CH, R¹⁶ is hydrogen, R¹⁷ and R¹⁸ are taken together to forman oxo (═O) group and R¹⁹ and R²⁰ are each hydrogen or methyl; and R² is

where each R¹³ is, independently of the other, selected from loweralkyl, methyl, hydroxy, lower alkoxy and methoxy.

When R² is lower alkyl in the compounds described herein, such as, forexample, the compounds of structural formulae (I)-(V), in someembodiments, R⁴ is

where D is as previously defined for structural formula (I).

When R² is

in the compounds described herein, such as, for example, the compoundsof structural formulae (I)-(V), in some embodiments, R¹¹ is selectedfrom the group consisting of hydroxy, methoxy, ethoxy, —NHCH₃,—NHCH₂CH₂OH, —NHCH₂CH(OH)CH₂OH, —NHCH₂CH(OH)(CH₃)₂, —N(CH₃)CH₂CH₂OH and—N(CH₃)C(CH₃)₂CH₂OH and Y is as previously defined.

When R² is

in the compounds described herein, such as the compounds of structuralformulae (I)-(V), in some embodiments the ring is connected to theremainder of the molecule at the 5-position

In other embodiments, it is connected to the remainder of the moleculeat the 6-position

In some embodiments, R¹⁶ is selected from lower n-alkanyl, lowerbranched alkanyl, lower cycloalkanyl and lower cycloalkanylmethyl. Insome embodiments, R⁴ is selected from

When R² is

in the compounds described herein, such as, for example, the compound ofstructural formulae (I)-(V), in some embodiments, R⁴ is selected fromlower cycloalkyl and lower cycloheteroalkyl optionally substituted atone or more ring carbon or heteroatoms with an R^(a) or an R^(b) group.In a specific embodiment, R⁴ is selected from cyclopropyl, cyclobutyl,cyclopentyl, cyclohexyl,

where R^(e) and R^(f) are selected from (C1-C3) alkanyl and methyl andR^(g) is benzyl. In some embodiments, each R^(e) is methyl. In someembodiments, R^(f) is ethyl.

When R⁴ is selected from lower alkyl, isopropyl, t-butyl, lowercycloalkyl,

in the compounds described herein, such as, for example, the compoundsof structural formulae (I)-(V), in some embodiments R² is selected from

where R⁸, R⁹, R¹⁰ and R¹³ are as previously defined for structuralformula (I). In a specific embodiment, R² is selected from any of theabove-described embodiments of these substituted phenyls. In otherembodiments, R² is

where R⁸ and R⁹ are a previously defined for structural formula (I).

When R⁴ is

where R¹⁵ is lower branched alkyl or t-butyl, and R² is

in the compounds described herein, such as, for example, the compoundsof structural formulae (I)-(V), in some embodiments at least one of R⁸or R¹⁰ is other than hydrogen. In other embodiments, at least two of R⁸,R⁹ and R¹⁰ are other than hydrogen. In still other embodiments, at leasttwo of R⁸, R⁹ and R¹³ are other than hydrogen.

In still other embodiments, either: (i) R⁹ is

and R¹⁰ is other than

or (ii) R¹⁰ is

and R⁹ is other than

In a specific embodiment of alternative (i), R¹⁰ is hydrogen. In aspecific embodiment of alternative (ii), R⁹ is hydrogen.

In still other embodiments, when R² is

where R⁸ and R⁹ are each hydrogen, then R¹⁰ is other than lower branchedalkyl, t-butyl or —O(CH₂)_(n)R^(b), where n is as previously defined forstructural formula (I) and R^(b) is selected from —NR^(c)R^(c),—C(O)OR^(a), —C(O)NR^(c)R^(c) and —C(O)NR^(a)R^(d).

In still other embodiments, when R² is

where R⁹ is hydrogen and R¹³ is selected from hydrogen, lower alkyl andmethyl, then R⁸ is other than —O(CH₂)_(n)R^(b), where n is as previouslydefined for structural formula (I) and R^(b) is —NR^(c)R^(c). In stillother embodiments, the compound is not any compound described in WO01/64656, WO 03/026665 or WO 03/026666, the disclosures of which areincorporated herein by reference.

When R⁴ is —(CH₂)_(n)—R^(b) in the compounds described herein, such as,for example, the compounds of structural formulae (II)-(V), in someembodiments R^(b) is selected from the group consisting of —OR^(a),—NR^(c)R^(c), —C(O)R^(a) and —C(O)NR^(c)R^(c), where each R^(c) isindependently selected from hydrogen and lower alkyl.

When R⁴ is

in the compounds described herein, such as, for example, the compoundsof structural formulae (I)-(V), in some embodiments R² is

where R⁹ is selected from the group consisting of —OR^(a), methoxy,isopropoxy, —OCH₂C(O)OR^(a), —OCH₂C(O)NHR^(a), —OCH₂C(O)NR^(a)R^(a) and—OCH₂CH₂NR^(a)R^(a); and R⁸ and R¹⁰ are as previously defined forstructural formula (I). In a specific embodiment, R⁸ and R¹⁰ areselected from one of the following combinations:

-   -   R⁸ and R¹⁰ are each the same lower alkyl or methyl;    -   R⁸ is lower alkyl or methyl and R¹⁰ is halo, flouro or chloro;        and    -   R⁸ and R¹⁰ are each the same halo, fluoro or chloro.

When R⁴ is selected from lower alkyl optionally monosubstituted with anR^(b) group, a lower cycloalkyl optionally monosubstituted with an R^(b)group and —C(R^(a)R^(a))_(n)—R^(b), where, R^(a) and R^(b) are aspreviously defined for structural formula (I), and/or L² is a loweralkylene linker in the compounds described herein, in some embodiments,R² is other than mono-substituted phenyl, 3-hydroxyphenyl, 3-halophenyl,3-chlorophenyl, 3-bromophenyl, 4-halophenyl, 4-chlorophenyl,4-bromophenyl, 3,4-dihalophenyl, 3,4-dichlorophenyl or3,4-dichlorophenyl. In a specific embodiment, R² is other than thesedefined groups in compounds in which R⁵ is —CF₃. In another specificembodiment, the compound is not any compound described in US2003/0171359 and/or WO 03/032997, the disclosures of which areincorporated herein by reference.

When R⁴ is

and R² is

in the compounds described herein, such as, for example, the compoundsof structural formulae (I)-(V), in some embodiments, R⁹ and R⁹ arenon-bulky substitutents. In a specific embodiment, R⁹ is other than

and R¹⁰ is other than

In another specific embodiment, R⁸, R⁹ and R¹⁰ are each, independentlyof one another, selected from hydrogen lower alkyl, methyl hydroxy,lower alkoxy, methoxy, halo, fluoro and chloro. In another specificembodiment, R⁸ is selected from hydrogen, lower alkyl, methyl, loweralkoxy and methoxy, R⁹ is selected from hydrogen, lower alkoxy andmethoxy and R¹⁰ is selected from lower alkyl, methyl, lower alkoxy,methoxy, halo, fluoro and

In all of the compounds described herein in which R⁴ is

in some embodiments D is selected from the group consisting of —CH₂—,—CF₂—, —CH₂CH₂—, —CF₂—CF₂— and —CH₂—CH₂—CH₂—.

In all of the compounds described herein having lower alkylsubstitutents or substituents including lower alkyl groups (e.g., loweralkoxy groups, etc.), in some embodiments the lower alkyl substituent orgroup is a saturated straight-chained, branched or cyclic alkyl (i.e.,an alkanyl).

Additional exemplary embodiments of the compounds described herein areillustrated in the following TABLES 1-14, below.

TABLE 1A Type A

Type B

No. Type n R²¹ R²² R²³ R^(4′) A549 H1299 101 A 1 H

H H + + 102 A 1 H H

H + + 103 A 1 H H

H + + 104 A 1 H

H H + + 105 A 1 H

H H + + 106 A 1 H H

H + + 107 A 2 H

H H + + 108 A 2 H

H H + + 109 A 2 H H

H + + 110 A 2 H H

Me + + 111 A 2 H Cl Cl H + + 112 A 2 H OMe Cl H + + 113 A 2 H H

H + + 114 A 2 H Cl Cl H + + 115 A 2 H H

H + + 116 A 2 Me H Me H + + 117 A 2 OMe H OMe H + + 118 A 2 OMe H CF₃ H− + 119 A 2 H H

H + + 120 A 2 H H

H + + 121 A 2 H

H H + + 122 A 2 H

H H + + 123 A 2 H

H Me + + 124 A 2 H

H Me + + 125 A 2 H

H Me + + 126 A 2 H

H Me + + 127 A 2 H H

Me + + 128 A 2 H H

Me + + 129 A 2 H H

Me + + 130 A 2 H H

Me + + 131 A 2 H

H H + + 132 A 2 H

H H + + 133 A 3 H

H H + + 134 A 3 H

H H + + 135 A 3 H H

H + + 136 A 3 H H

H + + 137 A 3 H H

H + + 138 A 3 H H

H + + 139 A 4 H

H H + + 140 A 4 H

H H + + 141 A 4 H

H H + + 142 A 4 H

H H + + 143 A 4 H H

H + + 144 A 4 H H

H + + 145 A 4 H H

H + + 146 A 4 H H

H + + 147 A 3 H

H + + 148 A 3 H

H H + + 149 A 4 H

H H + + 150 A 4 H

H H + + 151 A 2 H H

Me + + 152 A 2 H

Cl H + + 153 A 4 H

Cl H + + 154 (HCl salt) A 2 H

H H + + 155 TsOH salt) A 4 H

H H + + 156 (HCl salt) A 4 H

H H + + 157 A 3 H

Cl H + + 158 A 2 H

Cl H + + 159 A 4 H

Cl H + + 160 A 2 H

Me H + + 161 A 3 H

Me H + + 162 A 4 H

Me H + + 163 A 2 H

CF₃ H + + 164 A 2 H

CF₃ H + + 165 A 4 H

CF₃ H + + 166 A 2 H

Me H + + 167 B 2 H

— H + + 168 B 3 H

— H + + 169 B 4 H

— H + + 170 A 2 H

CH₂OH H 171 B 2 Me

— H + + 172 B 3 Me

— H + + 173 B 4 Me

— H + + 174 A 2 H

CH₂OH H + + 175 A 3 H

H H + − 176 A 3 H

H H + + 177 A 3 H

H H + +

TABLE 1B Type A

Type B

No. Type R²¹ R²² R²³ R^(4′) A549 H1299 178 A H

H H + + 179 A H H

H + + 180 A H

Cl H + + 181 A H

Me H + + 182 A H

CF₃ H + + 183 B H

— H + + 184 B Me

— H + +

TABLE 1C Type A

Type B

No. Type R²¹ R²² R²³ R^(4′) A549 H1299 185 A H

H H + + 186 A H H

H + + 187 A H

Cl H + + 188 A H

Me H + + 189 A H

CF₃ H + + 190 B H

— H + +

TABLE 1D Type A

Type B

No. Type R²¹ R²² R²³ R^(4′) A549 H1299 191 A H

H H 192 A H

H H 193 A H

H H 194 A H

H H 195 A H H

H 196 A H H

H 197 A H H

H 198 A H H

H

TABLE 2

No. R¹⁵ R²¹ R²² R²³ A549 H1299 199 t-butyl H H

+ 200 t-butyl H H

+ + 201 t-butyl H H

+ + 202 t-butyl H

H + + 203 t-butyl H

H + + 204 t-butyl H

H + + 205 cyclopropyl H H

+ + 206 cyclopropyl H

H + + 207 cyclopropyl H H

+ + 208 cyclopropyl H H

209 cyclopropyl H

H + + 210 cyclopropyl H

H + − 211 cyclopropyl H H

+ + 212 cyclopropyl H

Cl + + 213 cyclopropyl H

Me + + 214 cyclopropyl H

CF₃ + +

TABLE 3 Type A

Type B

No. Type R⁴ R²¹ R²² R²³ A549 H1299 215 A i-propyl H

H + + 216 A i-propyl H

Cl + + 217 A i-propyl H

Me + + 218 A i-propyl H

CF₃ + + 219 A i-propyl H H

+ + 220 A t-butyl H

H + + 221 A t-butyl H H

+ + 222 A t-butyl H

Cl + + 223 A t-butyl H

Me + + 224 A t-butyl H

CF₃ + + 225 B i-propyl H

— + + 226 B t-butyl H

— + + 227 B i-propyl Me

— + +

TABLE 4

No. Z¹ Z² R¹⁶ R¹⁹ R²⁰ R²¹ R²² R²³ R^(4′) A549 H1299 228 CH O H Me(R) H HOMe Cl H 229 CH O H Me(S) H Me H Me H + + 230 CH O H Me(R) H Me H Me H− + 231 CH O H Me(R) H Cl OMe H H − − 232 CH O H Me(R) H Cl OMe H H + −236 CH O H H H H H

H + + 237 CH O H CH₂CH₂OH H H H

H + + 238 CH O H CH₂CH₂OH H H H

H + + 239 CH O H H H H H

H − + 240 CH O H Me(S) H H H

H + − 241 CH O H Me(R) H H H

H 242 CH O H Me(S) H H H

H 243 CH S H H H H H

H 244 CH S H H H OMe H OMe H 245 CH S H H H H OMe Cl H 246 CH S Me H H HH

H 247 CH S Me H H OMe H OMe H 248 CH O H H H H H C(S)NH₂ H 249 CH O HCH₂CH₂OH H H H C(S)NH₂ H 250 CH S(O)₂ H Me H OMe H OMe H + + 251 CHS(O)₂ H Me H Me H Me H + + 252 CH S(O)₂ H Me H OMe OMe OMe H 253 CH O HH H H H

H 254 CH O H H H H H OH H + 255 CH O Me H H H H OH H + + 256 CH O H Me HH H

H + 257 CH O H Me(S) H Cl OMe Cl H 258 CH O H Me(R) H Cl OMe Cl H 259 CHO H CH₂CH₂OH H OMe H OMe H 260 CH O H CH₂CH₂OH H Cl OMe H H 261 CH O HMe(S) H OMe H OMe H 262 CH O H Me(R) H OMe H OMe H 263 CH S(O)₂ H Me MeOMe H OMe H − − 264 CH S(O)₂ H Me Me Me H Me H − + 265 CH S(O)₂ H Me MeOMe OMe OMe H + + 266 CH S H Me H OMe H OMe 267 CH S H Me H Me H Me H268 CH S H Me H OMe OMe OMe H 269 CH S(O)₂ H Me Me H H

H 270 CH S(O)₂ H Me Me H OMe Cl H 271 CH S H Me H H H

H 272 CH S H Me H H OMe Cl H 273 CH S H H H H H OH H 274 CH S H Me H H HOH H 275 CH S(O)₂ H Me Me H H OH H 276 CH S(O)₂ H Me H H H OH H 277 CHS(O)₂ H Me H H H

H 278 CH S(O)₂ H Me H H OMe Cl H 279 CH S H H H OMe OMe OMe H 280 CH S HMe Me H OMe Cl H 281 CH S H Me Me OMe H OMe H 282 CH S H Me Me Me H Me H283 CH S H Me Me OMe OMe OMe H 284 CH S H Me Me H H OH H 285 CH S H MeMe H H

H 286 CH S(O)₂ H Me H H OMe F H 287 CH S(O)₂ H Me Me H OMe F H + + 288CH S H Me H H OMe F H + + 289 CH S H Me Me H OMe F H + + 290 CH S H H HH OMe F H + + 291 CH S H H H Me O Me H − + 292 CH S(O)₂ H H H Me O Me H293 CH S(O)₂ H H H OMe H OMe H 294 CH S(O)₂ H H H H OMe Cl H 295 CHS(O)₂ H H H H H

H 296 CH S(O)₂ H H H OMe OMe OMe H 297 CH S(O)₂ H H H H H OH H 298 CHS(O)₂ H H H H OMe F H 299 CH O Me H H H H

H + − 300 CH O H Me(S) H H H

H + 301 CH O H Me(R) H H H

H 302 CH O H Me(S) H Cl OMe H H 303 CH O H Me(R) H Cl OMe H H 304 CH O HMe Me OMe Me Me H 305 CH O H Me Me OMe H OMe H + 306 CH O H Me Me Cl MeCl H + + 307 CH O H Me Me Cl OMe Cl H + 308 CH O H Me Me Cl H Cl H + 309CH O H Me Me OMe H CF₃ H + 310 CH O H Me Me Me H Me H + 311 CH O H Me MeOMe H OMe H + 316 CH O H Me Me Me H Me H 318 CH O H Me Me H OMe Cl H 319CH O H Me Me Me Cl Me H 320 CH O H Me Me CH₂OH H CH₂OH H + + 321 CH O HMe Me Cl H OMe H + + 322 CH O H Me Me H OMe Cl H 323 CH O H Me Me OMe HOMe Me + + 324 CH O Me Me Me Me H Me Me + − 325 CH O Me Me Me H OMe ClMe + + 326 CH O Me Me Me OMe H OMe Me + + 327 CH O H Me Me H OMe ClMe + + 328 CH O Me Me Me Me H Me H 329 CH O Me Me Me OMe H OMe H 330 CHO H Me Me H C(O)NHMe Cl H 331 CH O H Me Me H S(O)₂NHMe OMe H 332 CH O HMe Me H H

H 333 CH O H Me Me C(O)OMe H

H − − 334 CH O H Me Me CF₃ H

H + − 335 N O H Me Me Me OMe Me H 338 CH O H Me Me OMe OMe OMe H + + 339CH O H Me Me Me

Me H + + 340 N O H Me Me OMe OMe OMe H 350 N O H Me Me Me Cl Me H 352 NO H Me Me Cl OH Cl H + + 353 N O Me Me Me OMe OMe OMe H 354 N O H Me MeOMe

OMe H + + 355 CH O H Me Me H H

H + + 356 CH O H Me Me H H OH H + − 357 N O H Me Me H H

H 358 CH O H Me Me Me OH Cl H + 359 CH O H Me Me Me OMe Cl H + + 360 N OH Me Me H H

H 362 N O H Me Me H OMe Cl H 363 N O H Me Me OMe H OMe H 364 N O H Me MeH Cl Cl H 365 (HCl salt) N O H Me Me H H

H 366 (bis HCl salt) N O H Me Me H H

H 367 (nitrate salt) N O H Me Me H H

H 368 (bis nitrate salt) N O H Me Me H H

H 369 (mesylate salt) N O H Me Me H H

H 370 N O H Me Me H H

H 371 N O H Me Me H H t-butyl H + + 372 N O H Me Me H H OH H 374 N O HMe Me H OMe F H − − 375 N O H Me Me H H Cl H − − 376 N O H Me Me Cl H ClH + + 377 CH O H Me Me Me OMe Cl H 378 N O H Me Me H OCF₃ Cl H − − 379 NO H Me Me Me OMe Cl H 380 N O H Me Me H H

H 381 N O H Me Me Me OH Cl H 382 N O H Me Me Me OMe Me H − − 383 N O HMe Me H H i-propyl H 385 CH O H Me Me Cl OEt Me H + − 386 N O Me Me MeMe OMe Cl H 387 N O H Me Me Cl OEt Me H − − 388 N O Me Me Me H H

H + + 389 CH O H Me Me Me

Cl H 390 N O Me Me Me H H

H 391 CH O H Me Me H H

H + + 392 N O Me Me Me H H

H 393 CH O H Me Me Me

Cl H 394 CH O Me Me Me H H

H + − 395 CH O H Me Me H H

Me − + 396 CH O H Me Me H H

H 397 CH O Me Me Me H H

Me + + 398 N O H Me Me Me

Cl H − − 399 N O H Me Me H H

H 400 N O H Me Me H H

H 401 CH O Me Me Me H H

Me + + 402 CH O Me Me Me Me OMe Me Me + − 403 CH O Me Me Me Me

Cl Me + − 404 N O Me Me Me Me OMe Me H 405 N O Me Me Me Me

Cl H + + 406 N O H Me Me H H

H 407 CH O H Me Me H H

H + + 408 N O H Me Me Me

Cl H 409 N O H Me Me Me

Me H 410 CH O H Me Me Me i-propoxy Cl H 411 N O H Me Me Me i-propoxy ClH 412 N O H Me Me Me

Me H 413 N O H Me Me Me

Cl H 414 N O H Me Me Me

Cl H − − 415 N O H Me Me Me

Me H 416 CH O H Me Me H H

H 417 N O H Me Me H C(O)NHMe Cl H 418 N O H H H H H

H 419 N O H H H H

H H 421 N O H H H H Cl Cl H 422 N O H H H H OMe Cl H 423 N O H H H ClOMe Cl H 424 N O H H H OMe H OMe H 425 N O H H H H OMe F H 426 N O H H HH OMe H H 427 N O H H H H OCF₃ H H 428 N O H H H H OEt H H 429 N O H H HH OBu H H 430 N O H H H H

H H 431 N O H H H H O-iPr H H 432 N O H Me Me H

H H 433 N O H H H H OMe OMe H 434 N O H Me Me H

H H 435 N O H H H H

H H + + 436 N O H Me Me H

H H + 437 N O H H H Me H Me M 438 N O H Me Me Me H Me H 439 N O H H H HH i-propyl H − − 440 N O H H H H Me Cl H − − 441 N O H H H CF₃ H OMe H −− 442 N O H H H Cl H Cl H − − 443 N O H H H H H Br H − − 444 N O H H H HH t-butyl H − − 445 N O H H H OMe OMe OMe H 446 N O H H H H F F H − −447 N O H H H Me OMe Me H 448 N O H H H Me OH Me H + + 449 N O H H H H H

H + + 450 N O H H H H H

H + + 451 N O H H H H

H H − − 452 N O H H H H H

H 453 N O H H H H H

H + + 454 N O H Me Me H H

H − − 455 N O H Me Me H H

H + + 456 N O H Me Me H

H H + + 457 N O H Me Me H H

H 458 N O H Me Me H H

H + + 459 N O H Me(S) H H OMe Cl H 460 N O H Me(S) H OMe OMe OMe H 461 NO H Me(S) H Me H Me H 462 CH O H Me(R) H H C(O)NH₂ H H — + 463 CH O HMe(S) H CH₂NHBOC H H H 464 CH O H Me(R) H CH₂NHBOC H H H 465 CH O HMe(S) H CH₂NH₂ H H H 466 CH O H Me(R) H CH₂NH₂ H H H 467 CH O H Me Me HH

H 468 CH O H Me(S) H H H

H 469 CH O H Me(R) H H H

N H 470 CH O H Me Me H

H H − − 471 CH O H Me(S) H H

H H + − 472 N O H Me Me H C(O)NH₂ H H + + 473 N→O O H Me Me OMe OMe OMeH + + ^(††)In TABLE 4, compounds having chirality at the carbon labeledwith an asterisk (*) that, through substituent R¹⁹, designate aspecified stereochemistry were synthesized and tested as thesubstantially pure enantiomer; compounds that do not designate aspecified stereochemistry at this carbon atom were synthesized and, iftested, were tested as the racemate.

TABLE 5

No. m R⁷ R²¹ R²² R²³ R^(4′) A549 H1299 474 1 H H

H H − 475 1 H H

H H − 476 0 F H H

H 477 1 H H hexoxy H H − 478 1 H H OEt H H + 479 1 H H butoxy H H − 4801 H H

H H − 481 1 H H H

H − 482 1 H H H OH H − 483 1 H H OE H H − 484 1 H H OMe OMe H 485 1 H HF Cl H − 486 1 H H t-butyl H H 487 1 H H F H H − 488 1 H H H F H 489 1 HH Et H H − 490 1 H H

H H − 491 1 H H H

H − + 492 1 H H H

H − 493 1 H H H

H −/+ + 494 1 H H H

H + + 495 1 H H H

H −/+ + 496 1 H H H

H + 497 1 H H H

H + 498 1 H H H

H − 499 1 H H H

H 500 1 H H H

H + 501 1 H H

H H − 502 1 H H

H H − 503 1 F H H

H −/+ 504 1 H H H

H + 505 1 H H Me

H 506 1 H OMe OMe OMe H + 507 1 H Cl OH Cl H + 508 1 H H H

H − 509 1 H H H

H − 510 1 H H H

H − 511 1 H H i-propyl H H − 512 1 H OMe H OMe H + + 513 0 F H H ClH + + 514 1 H H H CF₃ H − 515 1 H H H

H + + 516 1 H H H

H + 517 1 H H H

H 518 1 H H OMe

H 519 1 H H H

H 520 1 H H H

H + 521 1 H Me H OH H + − 522 1 H F H CF₃ H − − 523 1 H Me H CF₃ H − +524 1 H F H F H − − 525 1 H H OMe Cl H 526 1 H H OCF₂ Cl H + − 527 1 HMe H Me H + 528 1 H Me Cl Me H + 529 1 H CH₂OH H CH₂OH H + + 530 1 H ClH Cl H + + 531 1 H OMe H CF₃ H − + 532 1 F OMe H OMe H + + 533 1 F Me ClMe H − + 534 1 F CH₂OH H CH₂OH H + 535 1 F Cl H Cl H + + 536 1 F OMe HCF₃ H + + 537 1 F Me H Me H + + 538 1 F Me H CF₃ H + − 539 1 H Cl H OMeH + + 540 0 H OMe H OMe H 541 0 H OMe H CF₃ H + + 542 0 H Me H CF₃ H − −543 0 H Cl H Cl H + + 544 0 H Me H Me H − − 545 0 H H H

Me 546 1 H OMe H OMe Me + − 547 1 H Me H Me Me + + 548 1 H H H

Me 549 1 H H H

Me + + 550 1 H H H

H + + 551 1 H H H H + + 552 1 F H H

H + + 553 1 F H

H H + + 554 1 H H C(O)NHMe Cl H + + 555 1 H H C(O)NHMe Cl Me 556 1 H HS(O)₂NHMe OMe H 557 1 H H H

H 558 1 H C(O)OMeH

H H + + 559 1 H CF₃ H

H + + 560 0 F OMe OMe OMe H + + 561 0 F OMe H OMe H + + 562 0 F Me H MeH + + 563 0 F Cl OH Cl H + + 564 0 F Cl H Cl H + + 565 0 F Me Cl Me H −− 566 0 F Me OH Cl H + + 567 0 F OMe H CF₃ H + + 568 0 F Me H CF₃ H − +569 1 H Me

Me H + + 570 0 H Me

Me H + + 571 1 F H H

H + + 572 1 F H H

Me + + 573 1 H Me

Me Me + + 574 1 F Me

Me Me + + 575 1 F Me

Me H + + 576 1 H H H

H + 577 2 H H

H H 578 2 H H

H H 579 2 H H

H H 580 2 H H H

H − 581 2 H H H

H + 582 2 H H H

H 583 1 H Me OH Cl H 584 1 H Me OMe Me H 585 1 H Me OMe Cl H 586 1 H H H

H − 587 1 H H

H H +

TABLE 6

No. Y¹ Y² Z³ R¹⁷ R¹⁸ R¹⁹ R²⁰ R²¹ R²² R²³ R^(4′) A549 H1299 588 N O CH MeMe H H OMe H OMe H + + 589 N O CH H H H H H H

H + + 590 N O CH Me Me H H H H

H + + 591 N O CH H H H H H H

H + + 592 N O CH Me Me H H H H

H — — 593 N O CH H H H CH₂CH₂OH OMe H OMe H 594 N O CH H H H CH₂CH₂OH MeH Me H 595 N S CH H H H H H H

H 596 N O CH H H Me Me H H

H 597 N O CH H H Me Me Me OMe Cl H + + 598 N O CH H H Me Me OMe H OMe H599 N O N H H Me Me H H

H 600 N O N H H H H H H

H 601 N O N H H H H H

H H 602 N O N H H H H H H

H 603 N O N H H H H H

H H + + 604 N O N H H H H H

H H − + 605 N O N H H H H H H

H + + 606 N O N H H H H H H

H + + 607 N O N H H H H H H

H + + 608 N O N H H H H H

H H + + 609 N O N H H H H H H

H + + 610 N O N H H H H H H

H + + 611 N O N H H H H OMe OMe OMe H + + 612 N O N H H H H H OMe Cl H −− 613 N O N H H H H H H

H + + 614 N O N H H H H H H

H + + 615 N O N H H H H H

Cl H + + 616 N O N H H H H H

Me H + +

TABLE 7 Type A Type B

No. Type R^(4′) R⁴ Y R¹¹ A549 HTC116 H1299 617 A H

NH OEt − 618 A H

NH OEt + 619 A H

O OMe − 620 A H

O OMe 621 A H

NH OH − 622 A H

O OH − 623 A H

O OMe − 624 A H

O

− 625 A H

O

+ + 626 A H

O

−/+ + 627 A H

O NHMe − 628 A H

O NHMe − 629 A H

O NHMe + + 630 A H

O NH(CH₂)₂OH − 631 A H

O NH(CH₂)₂OH + + 632 A H

O

+ + 633 A H

O

+ + 634 A H

O

+ − 635 A H

O

+ 636 A H

O OMe + − 637 A H

NH OEt 638 A H

O OMe − 639 A H

O OH − 640 A H

O

− 641 A H

O OMe − 642 A H

O NHMe + 643 A H

O N(Me)₂ − 644 A H

O

+ + 645 A H

O

+ + 646 B H

NH OEt + + + 647 B H

NH OEt + + 648 B H

NH OEt + + 649 B H

NH NHMe + + 650 A H

O OMe 651 A H

O OMe − 652 A H

O OMe − 653 B H

NH NHMe + + 654 B H

NH NHMe + + 655 B H

NH NHMe + + 656 A H

O OMe − − 657 A H

O OH − − 658 A H

NH Me − + 659 B H

NH N(Me)CH₂CH₂OH + + + 660 B H

NH NHC(Me)₂CH₂OH + + + 661 B Me

O OMe − − 662 A Me

O OMe − + 663 B Me

NH NHMe + + 664 B H

NH OEt + + 665 B H

NH NHMe + + 666 B Me

NH OEt + + 667 B H CH₂CH₂OH NH OEt − − − 668 B H

NH NHMe − + 669 B H

NH NHMe + + 670 B H

NH NHMe − − 671 A Me

O OMe − − 672 A Me

O OMe − − 673 A H

O OMe 674 A H

O OMe 675 A H

O NHMe − 676 A H

O OMe + 677 A H

O OMe + − 678 B H

NH OEt − −

TABLE 8

No. R^(4′) R²¹ R²² R²³ R³¹ R³² R³³ A549 H1299 679 H Cl H H Cl H H 680 HMe Me H Me Me H 681 H H H Br H H Br 682 H Cl H H Cl H H 683 H Me Me H MeMe H 684 H H Cl H H Cl H 685 H H OEt H H OEt H 686 H H OMe H H OMe H 687H H H H H H H 688 H Me H H Me H H 689 H H Br H H Br H 690 H H H H H H H691 H H H Br H H Br 692 H Me H H Me H H 693 H H

H H H OH + 694 H H

H H H

+ + 695 H H H

H F F 696 H H H

H Cl H 697 H H H

H Cl Cl + + 698 H H H

H H OCF₃ 699 H H H

H OCF₃ Cl + 700 H H H Me H OCF₃ Cl 701 H H CF₃ H H CF₃ H − − 702 H H OMeH H OMe H + + 703 H H CF₃ F H CF₃ F − 704 H H OEt H H OEt H + + 705 H HH OCF₃ H H OCF₃ − 706 H H Cl CF₃ H Cl CF₃ − 707 H H H OEt H H OEt + 708H H H OMe H H OMe + 709 H H OMe OMe H OEt H − 710 H H OMe OMe H OMeOMe + 711 H H H OH H H OH + 712 H H OMe OMe H H OH − 713 H H OEt H H OMeOMe + + 714 H H H OH H OEt H − 715 H H H OH H OMe OMe + 716 H H Cl H HCl H − 717 H H H Cl H H Cl + 718 H H t-butyl H H t-butyl H − − 719 H H FCl H F Cl − − 720 H H F H H F H + − 721 H H Me H H Me H − − 722 H H Et HH Et H − 723 H H

H H

H + 724 H H H

H H

− + 725 H H OMe OH H OMe OH + 726 H H Me OH H Me OH 727 H H

H H

H 728 H H

H H H OH 729 H H

H H H OH − 730 H H OH H H OH H + 731 H H OH Me H OH Me − 732 H H H

H H OH + 733 H H

H H

H − 734 H H

H H

H − 735 H H

H H

H 736 H H

H H H Cl − 737 H H

H H

H − 738 H H

H H

H − 739 H H

H H

H − 740 H H OH Cl H OH Cl 741 H H

Cl H Cl 742 H H OH F H OH F 743 H OMe OMe OMe OMe OMe OMe − 744 H H

H H H OH + 745 H H i-propoxy H H i-propoxy H − 746 H H H OH H

H − 747 H H

H H t-Bu H − 748 H H H

H t-Bu H − 749 H H H

H t-Bu H − 750 H H H

H t-Bu H − 751 H H H

H i-propoxy H −/+ 752 H H H

H i-propoxy H − 753 H H H

H OMe OMe − 754 H H H

H OMe OMe − 755 H H H

H H OMe + 756 H H H

H Me OH 757 H H Me

H H OH 758 H H Me

H Me OH 759 H H Me

H Me

+ + 760 H OMe OMe OMe H H OH − 761 H H H

H H OH 762 H H H

H H OH − 763 H H H OH H H

− + 764 H H i-pr H H H OH − 765 H OMe H OMe H H OH + 766 H H H

OMe H OMe − + 767 H H H

OMe H OMe − 768 H H H OH OMe H OMe + + 769 H OMe H OMe H H

− − 770 H H H

H H CF₃ + 771 H OMe H OMe H H

+ 772 H H H

H OEt H + + 773 H H OMe

H H OH 774 H H H

H H OH 775 H H H

H H Cl 776 H H H

CF₃ H OMe 777 H H H

H OMe OH 778 H H H

H OMe CF₃ 779 H H H

H F CF₃ 780 H H H

H Me Cl 781 H H OCF₃ Cl H H OH + + 782 H Br H CF₃ H H OH 783 H H H

H OCF₃ H 784 H H H

H CF₃ H 785 H H H

H Cl CF₃ 786 H H H OH H H OCF₃ 787 H Cl OH Me H H OH 788 H H H

H OMe Cl + 789 H H H

H OMe F 790 H H H

H Me OMe + 791 H H H

H H + + 792 H H H

H H + + 793 H H H

H Me CF₃ 794 H H H

H F Me 795 H

H

H H OH 796 H OH H

H H OH 797 H OH H

H H OH + − 798 H

H

H H OH − − 799 H H H OH H OMe Cl 800 H H OMe Cl H OMe Cl + − 801 H Me OHCl H OMe Cl 802 H Me OMe Cl H OMe Cl − − 803 H H H

H O-iPr Cl 804 H H H

Me OMe Cl 805 H OMe H OMe H

Cl 806 H CF₃ H OMe H

Cl + + 807 H H H

H OMe Cl 808 H Cl H Cl H OMe Cl + + 809 H Me H Me H OMe Cl 810 H CF₃ HOMe H OMe Cl + − 811 H Me Me Me H OMe Cl + − 812 H OMe H OMe H HOCF₃ + + 813 H Me H Me H H OCF₃ − − 814 H OMe H OMe Me H Me + + 815 HOMe H CF₃ Me H Me + + 816 H Me H CF₃ Me H Me + +/− 817 H Me H Me CF₃ HOMe + + 818 H OMe H OMe CF₃ H OMe + + 819 H CF₃ H OMe CF₃ H OMe + + 820H Me H CF₃ CF₃ H OMe +/− + 821 H OMe OMe OMe CF₃ H OMe + + 822 H Me OHCl CF₃ H OMe + + 823 H Cl H Cl CF₃ H OMe + + 824 H CH₂OH H CH₂OH CF₃ HOMe + −/+ 825 H Me Cl Me CF₃ H OMe + + 826 H H H

H Cl OMe 827 H H H

H

Cl 828 H Cl OH Cl H OMe Cl + + 829 H Cl OH Cl H OCF₃ Cl + + 830 H H H

H

Cl 831 H Cl OMe Cl H OMe Cl − − 832 H Cl OMe Cl H OCF₃ Cl + − 833 H H H

Cl OMe Cl 834 H Cl OMe Cl H Cl Cl + + 835 H OMe H OMe H Cl Cl + + 836 HOMe H OMe H OMe Cl 837 H OMe H OMe H OCF₃ Cl 838 H Me H Me H Cl Cl + −839 H OMe H OMe OMe H OMe + 840 H OMe H OMe OMe H OMe + + 841 H Me H MeMe H Me + + 842 H CH₂OH H CH₂OH OMe H OMe + + 843 H CH₂OH H CH₂OH H OMeCl + + 844 H CH₂OH H CH₂OH H Cl Cl + + 845 H Me Me Me OMe H OMe + + 846H Me H Me OMe H OMe + + 847 H Cl H Cl OMe H OMe + 848 H Me H CF₃ OMe HOMe + 849 H OMe H CF₃ OMe H OMe + + 850 Me H H

OMe H OMe − − 851 Me Me H Me OMe H OMe + + 852 Me OMe H OMe OMe HOMe + + 853 Me H OMe Cl OMe H OMe − − 854 Me H Cl OMe OMe H OMe − − 855H Me H Me OMe H OMe + + 856 H OMe H OMe H OMe OMe + + 857 H H H

H OMe OMe 858 H H H

H OMe OMe + + 859 H H H

H OMe OMe + + 860 H H OMe Cl H OMe OMe 861 H H H

OMe H OMe + + 862 H H H

H OMe OMe + + 863 H H H H OMe OMe + + 864 Me Me H Me H OMe OMe + + 865Me OMe H OMe H OMe OMe + + 866 H H

H OMe H OMe + + 867 Me H H

H OMe OMe − + 868 Me H H

H OMe OMe + + 869 Me H

H H OMe OMe + + 870 Me H H

H OMe OMe − − 871 H H C(O)NHMe Cl H OMe OMe + + 872 H H C(O)NHMe Cl OMeH OMe + + 873 H H C(O)NHMe Cl H OMe Cl 874 H H C(O)NHMe Cl H H OH 875 MeH C(O)NHMe Cl H OMe OMe + + 876 H OMe H OMe H C(O)NHMe Cl 877 H Me H MeH C(O)NHMe Cl 878 H H H

H C(O)NHMe Cl 879 H H S(O)₂NHMe OMe H OMe OMe + + 880 H H S(O)₂NHMe OMeOMe H OMe + + 881 H H S(O)₂NHMe OMe H Cl CF₃ 882 H H S(O)₂NHMe OMe HOCF₃ Cl 883 H H S(O)₂NHMe OMe H H Cl 884 H OMe H OMe H S(O)₂NHMe OMe + +885 H H H

H H OH + + 886 H H H

H OMe OMe + + 887 H C(O)OMe H

H OMe OMe + + 888 H C(O)Me H

H H OH + + 889 H CF₃ H

H OMe OMe + + 890 H H H

H H OH + + 891 H

H

H

− 892 H CF₃ H

CF₃ H

+ + 893 H Cl OH Cl Cl OMe Cl 894 H H

Me H

Me + + 895 H OMe OMe OMe Me H Me + + 896 H Me Cl Cl Me H Me + + 897 HCH₂OH H CH₂OH Me H Me + + 898 H Cl H Cl Me H Me − − 899 H Cl Cl Cl Me HMe + + 900 H Me Cl Me Me H Me − − 901 H Me

Me H OMe Cl 902 H Me

Me OMe H OMe + + 903 Me OMe H OMe H OMe Cl + + 904 Me Me OMe Cl H OMe Cl− − 905 Me Me

Me H OMe Cl + + 906 Me H H

H OMe Cl + + 907 Me H Me H H OMe Cl − + 908 Me OMe H OMe H Cl OMe 909 MeMe H Me H Cl OMe + + 910 Me H H

H Cl OMe + − 911 Me OMe H OMe Me Cl Me 912 Me Me H Me Me Cl Me − − 913Me H H

Me Cl Me + + 914 Me H H

Me Cl Me − - 915 H Me H Me H

Cl 916 H OMe H OMe H

Cl 917 H H OMe Cl H

Cl + + 918 H Me Cl Me H

Cl 919 H H C(O)NHMe Cl H Cl OMe 920 H H C(O)NHMe Cl H Cl CF3 921 Me HC(O)NHMe Cl Me Cl Me + + 922 H Me H Me H

Cl 923 H OMe H OMe H

Cl 924 H H OMe Cl H

Cl 925 H Me Cl Me H

Cl 926 H Me H Me H

Cl 927 H OMe H OMe H

Cl 928 H OMe Cl H H

Cl 929 H Me Cl Me H

Cl 930 H H Cl OMe H

Cl 931 H Me

Me OMe H OMe + + 932 H H H

OMe OMe OMe + + 933 H H H

OH OH

+ + 934 H H

H H H OH + 935 H H H

H i-pr H − 936 H H

H H i-pr H − 937 H H

H H i-pr H + 938 H H F Cl H F Cl − 939 H H H OH H H

− 940 H H H

OMe H OMe + + 941 H Cl OH Cl Cl OH Cl − 942 H Me OH Cl Me OH Cl + + 943H Cl OH Me H H OH + 944 H H H OH Cl OH Me 945 H H OMe OMe H

Me + + 946 H Me OH Cl H H 947 H H H

Me OH Me − + 948 H H H

Me OH Me − 949 H H H

Me OH Me + + 950 H H H

Me OH Me + + 951 H H H t-Bu H H OH − 952 H H H

Cl OH Me − + 953 H H H t-Bu H OMe OMe − 954 H H H

Cl OH Me − 955 H H H t-Bu H H

− 956 H H H t-Bu H H

− 957 H H H

Cl OH Me + 958 H H H

H H t-Bu − − 959 H Me OH Cl H H t-Bu − 960 H H H

H H t-Bu − 961 H H H

H H t-Bu −/+ 962 H H H

H H t-Bu − − 963 H H H

H H t-Bu 964 H H H OEt H H i-pr − 965 H H H

H H i-pr − 966 H H H

H H i-pr + 967 H H H

H CH₂OH CH₂OH − 968 H H H i-pr H H i-pr − + 969 H H OMe OMe H H OH + +970 H H H

H H CH₂NH₂ + + 971 H H

H H

H − 972 H H

H H

H 973 H H

H H

H − 974 H H H OH H

H + + 975 H H

H H H OH 976 H H H

H H OH − 977 H H H

H i-propoxy H − 978 H H H

H H OH − 979 H H H

Me OH Me − + 980 H H H

Me OH Me + 981 H H H

Me OH Me + 982 H H H

Me OH Me − 983 H H H

Cl OH Me + 984 H H H

Cl OH Me + 985 H H H

Me OMe Me − 986 H H H

Me OMe Me − 987 H H H

Me OMe Me − 988 H H H

H

H + + 989 H H H

H

H + + 990 H H H

H

H + + 991 H H H

Me OH Cl + 992 H H H

H

H − − 993 H H H

H

H + + 994 H H H

H H

995 H H H

H H

+ + 996 H H OH CF₃ H OH CF₃ −/+ 997 H H H Me H H Me + 998 H H Me H H MeH − 999 H H H

Cl OH Cl − 1000 H H H

Cl OH Cl 1001 H H H

Cl OH Cl − − 1002 H H H

H

H − 1003 H H H

H

H − 1004 H H H

H OH H − 1005 H H H

H OMe CH₂OH 1006 H H H

H Cl Cl + + 1007 Me Me H Me H Cl Cl − − 1008 Me H OMe Cl H Cl Cl 1009 MeOMe H OMe H Cl Cl − − 1010 Me Cl OMe Cl H Cl Cl 1011 Me H OCF₃ Cl H ClCl 1012 H H CH₂NHBoc H H CH₂NHBoc H + + 1013 H H CH₂NH₂ H H CH₂NH₂ H + +1014 H H

H H Cl Cl 1015 H H H CH₂NHBoc H H CH₂NHBoc + + 1016 H H H CH₂NHBoc H HCH₂NHBoc + + 1017 H H H CH₂NHBoc H Cl Cl 1018 H H H CH₂NHBoc H Cl Cl1019 H H

H H Cl Cl 1020 Me H

H H Cl Cl 1021 H H

H H OMe Cl + + 1022 H H C(O)NH₂ H H OMe Cl + + 1023 H H CH₂NHBoc H H OMeCl + + 1024 H H CH₂NHBoc H H OMe Cl + + 1025 H H

H H OMe Cl − + 1026 H H H OH H C(O)NH₂ H − + 1027 H Cl OH Cl H C(O)NH₂H + + 1028 H H OMe Cl H C(O)NH₂ H + + 1029 H H C(O)NH₂ H H C(O)NH₂ H − −1030 Me H C(O)NH₂ H H Cl C + + 1031 Me H

H H Cl Cl + + 1032 H H C(O)NH₂ H H Cl OMe − − 1033 H H C(O)NH₂ Cl H OMeCl + + 1034 H H C(O)NH₂ Cl H Cl OMe − + 1035 H H

H H Cl OMe + + 1036 H H C(O)NH₂ H H H H + +

TABLE 9

No. R^(4′) R^(2′) R² A549 H1299 1037 H H CH₂CH═CH₂ − 1038 Me H Me + +1039 Me Me Me + + 1040 Me H CH₂CH₂OH + + 1041 Me H i-propyl + + 1042 MeCH₂CH₂OH CH₂CH₂OH − + 1043 Me H CH₂CH═CH₂ + + 1044 H H Me − −

TABLE 10

No. R^(4′) R⁴⁰ R⁴¹ R⁴² A549 H1299 1045 H H OMe OMe − 1046 H H OMe Me + +1047 H OMe H Me + + 1048 H Me H F + + 1049 H OMe Cl OMe + +

TABLE 11

No. R²¹ R²² R²³ A549 H1299 1050 Me OMe Me + + 1051 H OMe F + + 1052 Me HMe + + 1053 OMe H OMe + + 1054 H H

+ + 1055 H H

− − 1056 H

H − − 1057 H

H − − 1058 H

H − − 1059 H H

− −

TABLE 12

No. R^(13a) R^(13b) R¹⁹ R²⁰ A549 H1299 1060 Me OH Me Me + + 1061 Me OMeMe Me + + 1062 Me OMe H H + + 1063 Me OH H H + + 1064 Me OH Me Me + +1065 Me OMe Me Me + −

TABLE 13 Type A

Type B

Type C

Type D

No. Type R¹⁶ R³¹ R³² R³³ A549 H1299 1066 B ethyl H OMe Cl 1067 B ethyl HCl Cl 1068 B ethyl — — — 1069 B ethyl H F OMe 1070 A ethyl H OMe Cl 1071A ethyl H Cl Cl 1072 C ethyl — — — 1073 A ethyl H F OMe 1074 B n-propylH OMe Cl + + 1075 B n-propyl H Cl Cl − − 1076 D n-propyl — — — + + 1077A n-propyl H F OMe + + 1078 B n-propyl H OMe Cl + + 1079 B n-propyl H ClCl + + 1080 C n-propyl — — — + + 1081 A n-propyl H F OMe + + 1082 Bn-butyl H OMe Cl + − 1083 B n-butyl H Cl Cl + + 1084 D n-butyl — — — + +1085 B n-butyl H F OMe + + 1086 A n-butyl H OMe Cl + + 1087 A n-butyl HCl Cl + + 1088 C n-butyl — — — + + 1089 A n-butyl H F OMe + + 1090 B

H OMe Cl + + 1091 B

H Cl Cl − − 1092 D

— — — + + 1093 B

H F OMe + + 1094 A

H OMe Cl + + 1095 A

H Cl Cl + + 1096 C

— — — + + 1097 A

H F OMe − + 1098 B

H OMe Cl − + 1099 B

H Cl Cl + + 1100 D

— — — + + 1101 B

H F OMe + + 1102 A

H OMe Cl + + 1103 A

H Cl Cl ° − 1104 C

— — — + + 1105 A

H F OMe + + 1106 B

H OMe Cl + + 1107 B

H Cl Cl − − 1108 D

— — — + + 1109 B

H F OMe + + 1110 A

H OMe Cl + + 1111 A

H Cl Cl + + 1112 C

— — — + + 1113 A

H F OMe + + 1114 B

H OMe Cl 1115 B

H Cl Cl + + 1116 D

— — — 1117 B

H F OMe + + 1118 A

H OMe Cl + + 1119 A

H Cl Cl + + 1120 C

— — — 1121 A

H F OMe + + 1122 C methyl — — — 1123 C methyl — — — 1124 C methyl — — —1125 C methyl — — — 1126 C methyl — — — 1127 B i-propyl H OMe Cl 1128 Bi-propyl H Cl Cl 1129 D i-propyl — — — 1130 B i-propyl H F OMe 1131 Ai-propyl H OMe Cl 1132 A i-propyl H Cl Cl 1133 C i-propyl — — — 1134 Cmethyl — — — 1135 C H — — — 1136 C H — — — 1137 C ethyl — — — 1138 Ci-propyl — — —

TABLE 14 No. Structure A549 H1299 1139

+ − 1140

− − 1141

− − 1142

+ − 1143

1144

1145

+ + 1146

+ − 1147

+ + 1148

1149

1150

1151

1152

1153

1154

1155

1156

1157

1158

+ + 1159

+ + 1160

+ + 1161

+ 1162

+ + 1163

+ + 1164

+ +

Those of skill in the art will appreciate that the 2,4-pyrimidinediaminecompounds described herein may include functional groups that can bemasked with progroups to create prodrugs. Such prodrugs are usually, butneed not be, pharmacologically inactive until converted into theiractive drug form. For example, ester groups commonly undergoacid-catalyzed hydrolysis to yield the parent carboxylic acid whenexposed to the acidic conditions of the stomach, or base-catalyzedhydrolysis when exposed to the basic conditions of the intestine orblood. Thus, when administered to a subject orally,2,4-pyrimidinediamines that include ester moieties may be consideredprodrugs of their corresponding carboxylic acid, regardless of whetherthe ester form is pharmacologically active.

In the prodrugs of the invention, any available functional moiety may bemasked with a progroup to yield a prodrug. Functional groups within the2,4-pyrimidinediamine compounds that may be masked with progroups forinclusion in a promoiety include, but are not limited to, amines(primary and secondary), hydroxyls, sulfanyls (thiols), carboxyls, etc.Myriad progroups suitable for masking such functional groups to yieldpromoieties that are cleavable under the desired conditions of use areknown in the art. All of these progroups, alone or in combinations, maybe included in the prodrugs of the invention.

In one illustrative embodiment, the prodrugs are compounds according tostructural formulae (I)-(VI) in which R^(a), R^(b) and R^(c) may be, inaddition to their previously-defined alternatives, a progroup.

In another illustrative embodiment, the prodrugs are compounds accordingto structural formulae (I)-(VI) in which R^(2′) and R^(4′) are each,independently of one another, a progroup. Specific examples of progroupsaccording to this embodiment of the invention include, but are notlimited to, —C(O)CH₃, —C(O)NHR^(h) and —S(O)₂R^(h), where R^(h) isselected from the group consisting of lower alkyl, (C5-C15) aryl and(C3-C8) cycloalkyl.

Those of skill in the art will appreciate that many of the compounds andprodrugs described herein, as well as the various compound speciesspecifically described and/or illustrated herein, may exhibit thephenomena of tautomerism, conformational isomerism, geometric isomerismand/or optical isomerism. For example, the compounds and prodrugs mayinclude one or more chiral centers and/or double bonds and as aconsequence may exist as stereoisomers, such as double-bond isomers(i.e., geometric isomers), enantiomers and diasteromers and mixturesthereof, such as racemic mixtures. As another example, the compounds andprodrugs may exist in several tautomeric forms, including the enol form,the keto form and mixtures thereof. As the various compound names,formulae and compound drawings within the specification and claims canrepresent only one of the possible tautomeric, conformational isomeric,optical isomeric or geometric isomeric forms, it should be understoodthat the invention encompasses any tautomeric, conformational isomeric,optical isomeric and/or geometric isomeric forms of the compounds orprodrugs having one or more of the utilities described herein, as wellas mixtures of these various different isomeric forms. In cases oflimited rotation around the 2,4-pryimidinediamine core structure, atropisomers are also possible and are also specifically included in thecompounds and/or prodrugs of the invention.

Depending upon the nature of the various substituents, the2,4-pyrimidinediamine compounds and prodrugs may be in the form ofsalts. Such salts include salts suitable for pharmaceutical uses(“pharmaceutically-acceptable salts”), salts suitable for veterinaryuses, etc. Such salts may be derived from acids or bases, as iswell-known in the art.

In some embodiments, the salt is a pharmaceutically acceptable salt.Generally, pharmaceutically acceptable salts are those salts that retainsubstantially one or more of the desired pharmacological activities ofthe parent compound and which are suitable for administration to humans.Pharmaceutically acceptable salts include acid addition salts formedwith inorganic acids or organic acids. Inorganic acids suitable forforming pharmaceutically acceptable acid addition salts include, by wayof example and not limitation, hydrohalide acids (e.g., hydrochloricacid, hydrobromic acid, hydriodic, etc.), sulfuric acid, nitric acid,phosphoric acid, and the like. Organic acids suitable for formingpharmaceutically acceptable acid addition salts include, by way ofexample and not limitation, acetic acid, trifluoroacetic acid, propionicacid, hexanoic acid, cyclopentanepropionic acid, glycolic acid, oxalicacid, pyruvic acid, lactic acid, malonic acid, succinic acid, malicacid, maleic acid, fumaric acid, tartaric acid, citric acid, palmiticacid, benzoic acid, 3-(4-hydroxybenzoyl) benzoic acid, cinnamic acid,mandelic acid, alkylsulfonic acids (e.g., methanesulfonic acid,ethanesulfonic acid, 1,2-ethane-disulfonic acid, 2-hydroxyethanesulfonicacid, etc.), arylsulfonic acids (e.g., benzenesulfonic acid,4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid,4-toluenesulfonic acid, camphorsulfonic acid, etc.),4-methylbicyclo[2.2.2]-oct-2-ene-1-carboxylic acid, glucoheptonic acid,3-phenylpropionic acid, trimethylacetic acid, tertiary butylacetic acid,lauryl sulfuric acid, gluconic acid, glutamic acid, hydroxynaphthoicacid, salicylic acid, stearic acid, muconic acid, and the like.

Pharmaceutically acceptable salts also include salts formed when anacidic proton present in the parent compound is either replaced by ametal ion (e.g., an alkali metal ion, an alkaline earth metal ion or analuminum ion) or coordinates with an organic base (e.g., ethanolamine,diethanolamine, triethanolamine, N-methylglucamine, morpholine,piperidine, dimethylamine, diethylamine, etc.).

The 2,4-pyrimidinediamine compounds and prodrugs, as well as the saltsthereof, may also be in the form of hydrates, solvates and N-oxides, asare well-known in the art.

5.3 Methods of Synthesis

The 2,4-pyrimidinediamine compounds and prodrugs may be synthesized viaa variety of different synthetic routes using commercially availablestarting materials and/or starting materials prepared by conventionalsynthetic methods. Suitable exemplary methods that may be routinelyadapted to synthesize the 2,4-pyrimidinediamine compounds and prodrugsare found in U.S. Pat. No. 5,958,935, the disclosure of which isincorporated herein by reference. Specific examples describing thesynthesis of numerous 2,4-pyrimidinediamine compounds and prodrugs, aswell as intermediates therefor, are described in copending U.S.application Ser. No. 10/355,543, filed Jan. 31, 2003 (US 2004-0029902published Feb. 12, 2004), WO 03/63794, copending U.S. application Ser.No. 10/631,029, filed Jul. 29, 2003, WO 2004/014312, copendingapplication Ser. No. 10/903,263, filed Jul. 30, 2004 and internationalapplication Ser. No. PCT/US04/024716, filed Jul. 30, 2004, the contentsof which are incorporated herein by reference. All of the compoundsdescribed herein (including prodrugs) can be prepared according to, orby routine adaptation of, these various methods.

A variety of exemplary synthetic routes that can be used to synthesizethe 2,4-pyrimidinediamine compounds and prodrugs are described inSchemes (I)-(XI), below. In Schemes (I)-(XI), like-numbered compoundshave similar structures. These methods may be routinely adapted tosynthesize the corresponding N2- and/or N4-alkylated compounds andprodrugs, as illustrated in Scheme (XII).

In one exemplary embodiment, the compounds can be synthesized fromsubstituted or unsubstituted uracils or thiouracils as illustrated inScheme (I), below:

In Scheme (I), R², R⁴, R⁵, R⁶, L¹ and L² are as previously defined forstructural formulae (I), X is a halogen (e.g., F, Cl, Br or I) and G andG′ are each, independently of one another, selected from the groupconsisting of O and S. Referring to Scheme (I), uracil or thiouracil 2is dihalogenated at the 2- and 4-positions using standard halogenatingagent POX₃ (or other standard halogenating agent) under standardconditions to yield 2,4-bishalo pyrimidine 4. Depending upon the R⁵substituent, in pyrimidine 4, the halide at the C4 position is morereactive towards nucleophiles than the halide at the C2 position. Thisdifferential reactivity can be exploited to synthesize2,4-pyrimidinediamines according structural formulae (I) by firstreacting 2,4-bishalopyrimidine 4 with one equivalent of amine 10,yielding 4N-substituted-2-halo-4-pyrimidineamine 8, followed by amine 6to yield a 2,4-pyrimidinediamine according structural formulae (I).2N,4N-bis(substituted)-2,4-pyrimidinediamines 12 and 14 can be obtainedby reacting 2,4-bishalopyrimidine 4 with excess 6 or 10, respectively.

In most situations, the C4 halide is more reactive towards nucleophiles,as illustrated in the Scheme. However, as will be recognized by skilledartisans, the identity of the R⁵ substituent may alter this reactivity.For example, when R⁵ is trifluoromethyl, a 50:50 mixture of4N-substituted-4-pyrimidineamine 8 and the corresponding2N-substituted-2-pyrimidineamine is obtained. Regardless of the identityof the R⁵ substituent, the regioselectivity of the reaction can becontrolled by adjusting the solvent and other synthetic conditions (suchas temperature), as is well-known in the art.

The reactions depicted in Scheme (I) may proceed more quickly when thereaction mixtures are heated via microwave. When heating in thisfashion, the following conditions may be used: heat to 175° C. inethanol for 5-20 min. in a Smith Reactor (Personal Chemistry) in asealed tube (at 20 bar pressure).

The uracil or thiouracil 2 starting materials may be purchased fromcommercial sources or prepared using standard techniques of organicchemistry. Commercially available uracils and thiouracils that can beused as starting materials in Scheme (I) include, by way of example andnot limitation, uracil (Aldrich #13,078-8; CAS Registry 66-22-8);2-thio-uracil (Aldrich #11,558-4; CAS Registry 141-90-2);2,4-dithiouracil (Aldrich #15,846-1; CAS Registry 2001-93-6);5-bromouracil (Aldrich #85,247-3; CAS Registry 51-20-7; 5-fluorouracil(Aldrich #85,847-1; CAS Registry 51-21-8); 5-iodouracil (Aldrich#85,785-8; CAS Registry 696-07-1); 5-nitrouracil (Aldrich #85,276-7; CASRegistry 611-08-5); 5-(trifluoromethyl)-uracil (Aldrich #22,327-1; CASRegistry 54-20-6). Additional 5-substituted uracils and/or thiouracilsare available from General Intermediates of Canada, Inc., Edmonton,Calif. (www.generalintermediates.com) and/or Interchim, Cedex, France(www.interchim.com), or may be prepared using standard techniques.Myriad textbook references teaching suitable synthetic methods areprovided infra.

Amines 6 and 10 may be purchased from commercial sources or,alternatively, may be synthesized utilizing standard techniques. Forexample, suitable amines may be synthesized from nitro precursors usingstandard chemistry. Specific exemplary reactions are provided in theExamples section. See also Vogel, 1989, Practical Organic Chemistry,Addison Wesley Longman, Ltd. and John Wiley & Sons, Inc.

Skilled artisans will recognize that in some instances, amines 6 and 10and/or substituents R⁵ and/or R⁶ on uracil or thiouracil 2 may includefunctional groups that require protection during synthesis. The exactidentity of any protecting group(s) used will depend upon the identityof the functional group being protected, and will be apparent to theseof skill in the art. Guidance for selecting appropriate protectinggroups, as well as synthetic strategies for their attachment andremoval, may be found, for example, in Greene & Wuts, Protective Groupsin Organic Synthesis, 3d Edition, John Wiley & Sons, Inc., New York(1999) and the references cited therein (hereinafter “Greene & Wuts”).

A specific embodiment of Scheme (I) utilizing 5-fluorouracil (Aldrich#32,937-1) as a starting material is illustrated in Scheme (II), below:

In Scheme (II), R², R⁴, L¹ and L² are as previously defined for Scheme(I). According to Scheme (II), 5-fluorouracil 3 is halogenated withPOCl₃ to yield 2,4-dichloro-5-fluoropyrimidine 5, which is then reactedwith excess amine 6 or 10 to yield N2,N4-bis substituted5-fluoro-2,4-pyrimidinediamine 11 or 13, respectively. Alternatively,non-bis-2N,4N-disubstituted-5-fluoro-2,4-pyrimidinediamine 9 may beobtained by reacting 2,4-dichloro-5-fluoropyrimidine 5 with oneequivalent of amine 10 (to yield2-chloro-N4-substituted-5-fluoro-4-pyrimidineamine 7) followed by one ormore equivalents of amine 6.

In another exemplary embodiment, the 2,4-pyrimidinediamine compounds ofthe invention may be synthesized from substituted or unsubstitutedcytosines as illustrated in Schemes (IIa) and (IIb), below:

In Schemes (IIa) and (IIb), R², R⁴, R⁵, R⁶, L¹, L² and X are aspreviously defined for Scheme (I) and PG represents a protecting group.Referring to Scheme (IIa), the C4 exocyclic amine of cytosine 20 isfirst protected with a suitable protecting group PG to yieldN4-protected cytosine 22. For specific guidance regarding protectinggroups useful in this context, see Vorbrüggen and Ruh-Pohlenz, 2001,Handbook of Nucleoside Synthesis, John Wiley & Sons, NY, pp. 1-631(“Vorbrüggen”). Protected cytosine 22 is halogenated at the C2 positionusing a standard halogenation reagent under standard conditions to yield2-chloro-4N-protected-4-pyrimidineamine 24. Reaction with amine 6followed by deprotection of the C4 exocyclic amine and reaction withamine 10 yields a 2,4-pyrimidinediamine according to structural formulae(I).

Alternatively, referring to Scheme (IIb), cytosine 20 may be reactedwith amine 10 or protected amine 21 to yield N4-substituted cytosine 23or 27, respectively. These substituted cytosines may then be halogenatedas previously described, deprotected (in the case of N4-substitutedcytosine 27) and reacted with amine 6 to yield a 2,4-pyrimidinediamineaccording to structural formulae (I).

Commercially-available cytosines that may be used as starting materialsin Schemes (IIa) and (IIb) include, but are not limited to, cytosine(Aldrich #14,201-8; CAS Registry 71-30-7); N⁴-acetylcytosine (Aldrich#37,791-0; CAS Registry 14631-20-0); 5-fluorocytosine (Aldrich#27,159-4; CAS Registry 2022-85-7); and 5-(trifluoromethyl)-cytosine.Other suitable cytosines useful as starting materials in Schemes (IIa)are available from General Intermediates of Canada, Inc., Edmonton,Calif. (www.generalintermediates.com) and/or Interchim, Cedex, France(www.interchim.com), or may be prepared using standard techniques.Myriad textbook references teaching suitable synthetic methods areprovided infra.

In still another exemplary embodiment, the 2,4-pyrimidinediaminecompounds may be synthesized from substituted or unsubstituted2-amino-4-pyrimidinols as illustrated in Scheme (III), below:

In Scheme (III), R², R⁴, R⁵, R⁶, L¹, L² and X are as previously definedfor Scheme (I) and Z is a leaving group as discussed in more detail inconnection with Scheme IV, infra. Referring to Scheme (III),2-amino-4-pyrimidinol 30 is reacted with amine 6 (or optionallyprotected amine 21) to yield N2-substituted-4-pyrimidinol 32, which isthen halogenated as previously described to yieldN2-substituted-4-halo-2-pyrimidineamine 34. Optional deprotection (forexample if protected amine 21 was used in the first step) followed byreaction with amine 10 affords a 2,4-pyrimidinediamine according tostructural formulae (I). Alternatively, pyrimidinol 30 can be reactedwith acylating agent 31.

Suitable commercially-available 2-amino-4-pyrimidinols 30 that can beused as starting materials in Scheme (III) are available from GeneralIntermediates of Canada, Inc., Edmonton, Calif.(www.generalintermediates.com) and/or Interchim, Cedex, France(www.interchim.com), or may be prepared using standard techniques.Myriad textbook references teaching suitable synthetic methods areprovided infra.

Alternatively, the 2,4-pyrimidinediamine compounds may be prepared fromsubstituted or unsubstituted 4-amino-2-pyrimidinols as illustrated inScheme (IV), below:

In Scheme (IV), R², R⁴, R⁵, R⁶, L¹ and L² are as previously defined forScheme (I) and Z represents a leaving group. Referring to Scheme (IV),the C2-hydroxyl of 4-amino-2-pyrimidinol 40 is more reactive towardsnucleophiles than the C4-amino such that reaction with amine 6 yieldsN2-substituted-2,4-pyrimidinediamine 42. Subsequent reaction withcompound 44, which includes a good leaving group Z, or amine 10 yields a2,4-pyrimidinediamine according to structural formulae (I). Compound 44may include virtually any leaving group that can be displaced by theC4-amino of N2-substituted-2,4-pyrimidinediamine 42. Suitable leavinggroups Z include, but are not limited to, halogens, methanesulfonyloxy(mesyloxy; “OMs”), trifluoromethanesulfonyloxy (“OTf”) andp-toluenesulfonyloxy (tosyloxy; “OTs”), benzene sulfonyloxy (“besylate”)and metanitro benzene sulfonyloxy (“nosylate”). Other suitable leavinggroups will be apparent to those of skill in the art.

Substituted 4-amino-2-pyrimidinol starting materials may be obtainedcommercially or synthesized using standard techniques. Myriad textbookreferences teaching suitable synthetic methods are provided infra.

In still another exemplary embodiment, the 2,4-pyrimidinediaminecompounds can be prepared from 2-chloro-4-aminopyrimidines or2-amino-4-chloropyrimidines as illustrated in Scheme (V), below:

In Scheme (V), R², R⁴, R⁵, R⁶, L¹, L² and X are as defined for Scheme(I) and Z is as defined for Scheme (IV). Referring to Scheme (V),2-amino-4-chloropyrimidine 50 is reacted with amino 10 to yield4N-substituted-2-pyrimidineamine 52 which, following reaction withcompound 31 or amine 6, yields a 2,4-pyrimidinediamine according tostructural formulae (I). Alternatively, 2-chloro-4-amino-pyrimidine 54may be reacted with compound 44 followed by amine 6 to yield a compoundaccording to structural formulae (I).

A variety of pyrimidines 50 and 54 suitable for use as startingmaterials in Scheme (V) are commercially available from GeneralIntermediates of Canada, Inc., Edmonton, Calif.(www.generalintermediates.com) and/or Interchim, Cedex, France(www.interchim.com), or may be prepared using standard techniques.Myriad textbook references teaching suitable synthetic methods areprovided infra.

Alternatively, 4-chloro-2-pyrimidineamines 50 may be prepared asillustrated in Scheme (Va):

In Scheme (Va), R⁵ and R⁶ are as previously defined for structuralformulae (I). In Scheme (Va), dicarbonyl 53 is reacted with guanidine toyield 2-pyrimidineamine 51. Reaction with peracids likem-chloroperbenzoic acid, trifluoroperacetic acid or urea hydrogenperoxide complex yields N-oxide 55, which is then halogenated to give4-chloro-2-pyrimidineamine 50. The corresponding4-halo-2-pyrimidineamines may be obtained by using suitable halogenationreagents.

In yet another exemplary embodiment, the 2,4-pyrimidinediamine compoundscan be prepared from substituted or unsubstituted uridines asillustrated in Scheme (VI), below:

In Scheme (VI), R², R⁴, R⁵, R⁶, L¹, L² and X are as previously definedfor Scheme (I) and the superscript PG represents a protecting group, asdiscussed in connection with Scheme (IIb). According to Scheme (VI),uridine 60 has a C4 reactive center such that reaction with amine 10 orprotected amine 21 yields N4-substituted cytidine 62 or 64,respectively. Acid-catalyzed deprotection of N4-substituted 62 or 64(when “PG” represents an acid-labile protecting group) yieldsN4-substituted cytosine 28, which may be subsequently halogenated at theC2-position and reacted with amine 6 to yield a 2,4-pyrimidinediamineaccording to structural formulae (I).

Cytidines may also be used as starting materials in an analogous manner,as illustrated in Scheme (VII), below:

In Scheme (VII), R², R⁴, R⁵, R⁶, L¹, L² and X are as previously definedin Scheme (I) and the superscript PG represents a protecting group asdiscussed above. Referring to Scheme (VII), like uridine 60, cytidine 70has a C4 reactive center such that reaction with amine 10 or protectedamine 21 yields N4-substituted cytidine 62 or 64, respectively. Thesecytidines 62 and 64 are then treated as previously described for Scheme(VI) to yield a 2,4-pyrimidinediamine according to structural formulae(I).

Although Schemes (VI) and (VII) are exemplified with ribosylnucleosides,skilled artisans will appreciate that the corresponding 2′-deoxyribo and2′,3′-dideoxyribo nucleosides, as well as nucleosides including sugarsor sugar analogs other than ribose, would also work.

Numerous uridines and cytidines useful as starting materials in Schemes(VI) and (VII) are known in the art, and include, by way of example andnot limitation, 5-trifluoromethyl-2′-deoxycytidine (Chem. Sources #ABCRF07669; CAS Registry 66,384-66-5); 5-bromouridine (Chem. Sources Int'l2000; CAS Registry 957-75-5); 5-iodo-2′-deoxyuridine (Aldrich #1-775-6;CAS Registry 54-42-2); 5-fluorouridine (Aldrich #32,937-1; CAS Registry316-46-1); 5-iodouridine (Aldrich #85,259-7; CAS Registry 1024-99-3);5-(trifluoromethyl)uridine (Chem. Sources Int'l 2000; CAS Registry70-00-8); 5-trifluoromethyl-2′-deoxyuridine (Chem. Sources Int'l 2000;CAS Registry 70-00-8). Additional uridines and cytidines that can beused as starting materials in Schemes (VI) and (VII) are available fromGeneral Intermediates of Canada, Inc., Edmonton, Calif.(www.generalintermediates.com) and/or Interchim, Cedex, France(www.interchim.com), or may be prepared using standard techniques.Myriad textbook references teaching suitable synthetic methods areprovided infra.

As will be recognized by skilled artisans, certain2,4-pyrimidinediamines compounds synthesized via the exemplary methodsdescribed above, or by other well-known means, may also be utilized asstarting materials and/or intermediates to synthesize additional2,4-pyrimidinediamine compounds. A specific example is illustrated inScheme (VIII), below:

In Scheme (VIII), R⁴, R⁵, R⁶, L², Y, R^(a) and R^(c) are as previouslydefined for structural formulae (I). Each R^(a′) is independently anR^(a), and may be the same or different from the illustrated R^(a).Referring to Scheme (VIII), carboxylic acid or ester 100 may beconverted to amide 104 by reaction with amine 102. In amine 102, R^(a′)may be the same or different than R^(a) of acid or ester 100. Similarly,carbonate ester 106 may be converted to carbamate 108.

A second specific example is illustrated in Scheme (IX), below:

In Scheme (IX) R⁴, R⁵, R⁶, L², Y and R^(c) are as previously defined forstructural formulae (I). Referring to Scheme (IX), amide 110 or 116 maybe converted to amine 114 or 118, respectively, by borane reduction withborane methylsulfide complex 112. Other suitable reactions forsynthesizing 2,4-pyrimidinediamine compounds from 2,4-pyrimidinediaminestarting materials will be apparent to those of skill in the art.

Compounds including alkyl groups at the amine groups at the 2- and/or4-position of the 2,4-pyrimidinediamine ring can be prepared usingroutine alkylation procedures. An exemplary scheme for selectivelymethylating the amine group at the 4-position of the2,4-pyrimidinediamine is illustrated in Scheme (XII):

In Scheme (XII), L¹, L², R², R⁴, R⁵ and R⁶ are as previously defined forstructural formula (I). The above Scheme can be routinely adapted tosynthesize other N-alkylated compounds.

Compounds that are entiomerically or diasteriomerically pure can besynthesized via chiral-specific syntheses utilizing enantiomerically ordiasteriomerically pure starting reagents as is known in the art.Alternatively, specified enantiomers, diastereomers and/or mixturesthereof can be isolated utilizing conventional chiral separationtechniques.

Although many of the synthetic schemes discussed above do not illustratethe use of protecting groups, skilled artisans will recognize that insome instances certain substituents, such as, for example, R² and/or R⁴,may include functional groups requiring protection. The exact identityof the protecting group used will depend upon, among other things, theidentity of the functional group being protected and the reactionconditions used in the particular synthetic scheme, and will be apparentto those of skill in the art. Guidance for selecting protecting groupsand chemistries for their attachment and removal suitable for aparticular application can be found, for example, in Greene & Wuts,supra.

Prodrugs as described herein may be prepared by routine modification ofthe above-described methods. Alternatively, such prodrugs may beprepared by reacting a suitably protected 2,4-pyrimidinediamine ofstructural formula (I), (II), (III), (IV) and/or (V) with a suitableprogroup. Conditions for carrying out such reactions and fordeprotecting the product to yield a prodrugs as described herein arewell-known.

Myriad references teaching methods useful for synthesizing pyrimidinesgenerally, as well as starting materials described in Schemes (I)-(IX),are known in the art. For specific guidance, the reader is referred toBrown, D. J., “The Pyrimidines”, in The Chemistry of HeterocyclicCompounds, Volume 16 (Weissberger, A., Ed.), 1962, IntersciencePublishers, (A Division of John Wiley & Sons), New York (“Brown I”);Brown, D. J., “The Pyrimidines”, in The Chemistry of HeterocyclicCompounds, Volume 16, Supplement I (Weissberger, A. and Taylor, E. C.,Ed.), 1970, Wiley-Interscience, (A Division of John Wiley & Sons), NewYork (Brown II″); Brown, D. J., “The Pyrimidines”, in The Chemistry ofHeterocyclic Compounds, Volume 16, Supplement II (Weissberger, A. andTaylor, E. C., Ed.), 1985, An Interscience Publication (John Wiley &Sons), New York (“Brown III”); Brown, D. J., “The Pyrimidines” in TheChemistry of Heterocyclic Compounds, Volume 52 (Weissberger, A. andTaylor, E. C., Ed.), 1994, John Wiley & Sons, Inc., New York, pp. 1-1509(Brown IV″); Kenner, G. W. and Todd, A., in Heterocyclic Compounds,Volume 6, (Elderfield, R. C., Ed.), 1957, John Wiley, New York, Chapter7 (pyrimidines); Paquette, L. A., Principles of Modern HeterocyclicChemistry, 1968, W. A. Benjamin, Inc., New York, pp. 1-401 (uracilsynthesis pp. 313, 315; pyrimidine synthesis pp. 313-316; aminopyrimidine synthesis pp. 315); Joule, J. A., Mills, K. and Smith, G. F.,Heterocyclic Chemistry, 3^(rd) Edition, 1995, Chapman and Hall, London,UK, pp. 1-516; Vorbrüggen, H. and Ruh-Pohlenz, C., Handbook ofNucleoside Synthesis, John Wiley & Sons, New York, 2001, pp. 1-631(protection of pyrimidines by acylation pp. 90-91; silylation ofpyrimidines pp. 91-93); Joule, J. A., Mills, K. and Smith, G. F.,Heterocyclic Chemistry, 4^(th) Edition, 2000, Blackwell Science, Ltd,Oxford, UK, pp. 1-589; and Comprehensive Organic Synthesis, Volumes 1-9(Trost, B. M. and Fleming, I., Ed.), 1991, Pergamon Press, Oxford, UK.

5.4 Activity of the Antiproliferative Compounds

Active 2,4-pyrimidinediamine compounds typically inhibit proliferationof desired cells, such as tumor cells, with an IC₅₀ in the range ofabout 1 mM or less, as measured in a standard in vitro cellularproliferation assay. Of course, skilled artisans will appreciate thatcompounds which exhibit lower IC₅₀s, for example on the order of 100 μM,20 μM, 10 μM, 1 μM, 100 nM, 10 nM, 1 nM, or even lower, may beparticularly useful in therapeutic applications. The antiprolierativeactivity may be cytostatic or it may be cytotoxic. In instances whereantiproliferative activity specific to a particular cell type isdesired, the compound may be assayed for activity with the desired celltype and counter-screened for a lack of activity against other celltypes. The desired degree of “inactivity” in such counter screens, orthe desired ratio of activity vs. inactivity may vary for differentsituations, and may be selected by the user.

5.5 Uses of the Antiproliferative Compounds

The antiproliferative 2,4-pyrimidinediamine compounds, including thevarious salts, prodrugs, hydrates and N-oxide forms thereof, may be usedto inhibit cell proliferation in a variety of contexts. According tosome embodiments of the method, a cell or population of cells iscontacted with an amount of such a compound effective to inhibitproliferation of the cell or cell population. The compound may actcytotoxically to kill the cell, or cytostatically to inhibitproliferation without killing the cell.

In some embodiments, the methods may be practiced as a therapeuticapproach towards the treatment of proliferative disorders. Thus, in aspecific embodiment, the 2,4-pyrimidinediamine compounds (and thevarious forms described herein) may be used to treat proliferativedisorders in animal subjects, including humans. The method generallycomprises administering to the subject an amount of a compound of theinvention, or a salt, prodrug, hydrate or N-oxide thereof, effective totreat the disorder. In one embodiment, the subject is a mammal,including, but not limited to, bovine, horse, feline, canine, rodent, orprimate. In another embodiment, the subject is a human.

A variety of cellular proliferative disorders may be treated with thecompounds of the present invention. In one embodiment, the compounds areused to treat various cancers in afflicted subjects. Cancers aretraditionally classified based on the tissue and cell type from whichthe cancer cells originate. Carcinomas are considered cancers arisingfrom epithelial cells while sarcomas are considered cancers arising fromconnective tissues or muscle. Other cancer types include leukemias,which arise from hematopoietic cells, and cancers of nervous systemcells, which arise from neural tissue. For non-invasive tumors, adenomasare considered benign epithelial tumors with glandular organizationwhile chondomas are benign tumor arising from cartilage. In the presentinvention, the described compounds may be used to treat proliferativedisorders encompassed by carcinomas, sarcomas, leukemias, neural celltumors, and non-invasive tumors.

In a specific embodiment, the compounds are used to treat solid tumorsarising from various tissue types, including, but not limited to,cancers of the bone, breast, respiratory tract (e.g., bladder), brainreproductive organs, digestive tract, urinary tract, eye, liver, skin,head, neck, thyroid, parathyroid, and mestastatic forms thereof.

Specific proliferative disorders include the following: a) proliferativedisorders of the breast include, but are not limited to, invasive ductalcarcinoma, invasive lobular carcinoma, ductal carcinoma, lobularcarcinoma in situ, and metastatic breast cancer; b) proliferativedisorders of the skin include, but are not limited to, basal cellcarcinoma, squamous cell carcinoma, malignant melanoma, and Karposi'ssarcoma; c) proliferative disorders of the respiratory tract include,but are not limited to, small cell and non-small cell lung carcinoma,bronchial adema, pleuropulmonary blastoma, and malignant mesothelioma;d) proliferative disorders of the brain include, but are not limited to,brain stem and hyptothalamic glioma, cerebellar and cerebralastrocytoma, medullablastoma, ependymal tumors, oligodendroglial,meningiomas, and neuroectodermal and pineal tumors; e) proliferativedisorders of the male reproductive organs include, but are not limitedto, prostate cancer, testicular cancer, and penile cancer f)proliferative disorders of the female reproductive organs include, butare not limited to, uterine cancer (endometrial), cervical, ovarian,vaginal, vulval cancers, uterine sarcoma, ovarian germ cell tumor; g)proliferative disorders of the digestive tract include, but are notlimited to, anal, colon, colorectal, esophageal, gallbladder, stomach(gastric), pancreatic cancer, pancreatic cancer-Islet cell, rectal,small-intestine, and salivary gland cancers; h) proliferative disordersof the liver include, but are not limited to, hepatocellular carcinoma,cholangiocarcinoma, mixed hepatocellular cholangiocarcinoma, and primaryliver cancer; i) proliferative disorders of the eye include, but are notlimited to, intraocular melanoma, retinoblastoma, and rhabdomyosarcoma;j) proliferative disorders of the head and cancers include, but are notlimited to, laryngeal, hypopharyngeal, nasopharyngeal, oropharyngealcancers, and lip and oral cancer, squamous neck cancer, metastaticparanasal sinus cancer; k) proliferative disorders of the lymphomasinclude, but are not limited to, various T cell and B cell lymphomas,non-Hodgkins lymphoma, cutaneous T cell lymphoma, Hodgkins disease, andlymphoma of the central nervous system; l) leukemias include, but arenot limited to, acute myeloid leukemia, acute lymphoblastic leukemia,chronic lymphocytic leukemia, chronic myelogenous leukemia, and haircell leukemia, m) proliferative disorders of the thyroid include thyroidcancer, thymoma, and malignant thymoma; n) proliferative disorders ofthe urinary tract include, but are not limited to, bladder cancer; o)sarcomas include, but are not limited to, sarcoma of the soft tissue,osteosarcoma, malignant fibrous histiocytoma, lymphosarcoma, andrhabdomyosarcoma.

It is to be understood that the descriptions of proliferative disordersis not limited to the conditions described above, but encompasses otherdisorders characterized by uncontrolled growth and malignancy. It isfurther understood that proliferative disorders include variousmetastatic forms of the tumor and cancer types described herein. Thecompounds of the present invention may be tested for effectivenessagainst the disorders described herein, and a therapeutically effectiveregimen established. Effectiveness, as further described below, includesreduction or remission of the tumor, decreases in the rate of cellproliferation, or cytostatic or cytotoxic effect on cell growth.

5.6 Combination Therapies

The compounds of the present invention may be used alone, in combinationwith one another, or as an adjunct to, or in conjunction with, otherestablished antiproliferative therapies. Thus, the compounds of thepresent invention may be used with traditional cancer therapies, such asionization radiation in the form of γ-rays and x-rays, deliveredexternally or internally by implantation of radioactive compounds, andas a follow-up to surgical removal of tumors.

In another aspect, the compounds of the present invention may be usedwith other chemotherapeutic agents useful for the disorder or conditionbeing treated. These compounds may be administered simultaneously,sequentially, by the same route of administration, or by a differentroute.

In one embodiment, the present compounds may be used with otheranti-cancer or cytotoxic agents. Various classes of anti-cancer andanti-neoplastic compounds include, but are not limited to, alkylatingagents, antimetabolites, vinca alkyloids, taxanes, antibiotics, enzymes,cytokines, platinum coordination complexes, substituted ureas, tyrosinekinase inhibitors, hormones and hormone antagonists. Exemplaryalkylating agents include, by way of example and not limitation,mechlorothamine, cyclophosphamide, ifosfamide, melphalan, chlorambucil,ethyleneimines, methylmelamines, alkyl sulfonates (e.g., busulfan), andcarmustine. Exemplary antimetabolites include, by way of example and notlimitation, folic acid analog methotrexate; pyrmidine analogfluorouracil, cytosine arbinoside; purine analogs mecaptopurine,thioguanine, and azathioprine. Exemplary vinca alkyloids include, by wayof example and not limitation, vinblastine, vincristine, paclitaxel, andcolchicine. Exemplary antibiotics include, by way of example and notlimitation, actinomycin D, daunorubicin, and bleomycin. An exemplaryenzyme effective as anti-neoplastic agents include L-asparaginase.Exemplary coordination compounds include, by way of example and notlimitation, cisplatin and carboplatin. Exemplary hormones and hormonerelated compounds include, by way of example and not limitation,adrenocorticosteroids prednisone and dexamethasone; aromatase inhibitorsamino glutethimide, formestane, and anastrozole; progestin compoundshydroxyprogesteron caproate, medroxyprogesterone; and anti-estrogencompound tamoxifen.

These and other useful anti-cancer compounds are described in MerckIndex, 13th Ed. (O'Neil M. J. et al., ed) Merck Publishing Group (2001)and Goodman and Gilmans The Pharmacological Basis of Therapeutics, 10thEdition, Hardman, J. G. and Limbird, L. E. eds., pg. 1381-1287, McGrawHill, (1996), both of which are incorporated by reference herein.

Additional anti-proliferative compounds useful in combination with thecompounds of the present invention include, by way of example and notlimitation, antibodies directed against growth factor receptors (e.g.,anti-Her2); antibodies for activating T cells (e.g., anti-CTLA-4antibodies); and cytokines such as interferon-α and interferon-γ,interleukin-2, and GM-CSF.

5.7 Formulations and Administration

When used to treat or prevent such diseases, the active compounds may beadministered singly, as mixtures of one or more active compounds or inmixture or combination with other agents useful for treating suchdiseases and/or the symptoms associated with such diseases. The activecompounds may also be administered in mixture or in combination withagents useful to treat other disorders or maladies, such as steroids,membrane stabilizers. The active compounds or prodrugs may beadministered per se, or as pharmaceutical compositions, comprising anactive compound or prodrug.

Pharmaceutical compositions comprising the active compounds of theinvention (or prodrugs thereof) may be manufactured by means ofconventional mixing, dissolving, granulating, dragee-making levigating,emulsifying, encapsulating, entrapping or lyophilization processes. Thecompositions may be formulated in conventional manner using one or morephysiologically acceptable carriers, diluents, excipients or auxiliarieswhich facilitate processing of the active compounds into preparationswhich can be used pharmaceutically (see Remingtons's PharmaceuticalSciences, 15^(th) Ed., Hoover, J. E. ed., Mack Publishing Co. (2003)

The active compound or prodrug may be formulated in the pharmaceuticalcompositions per se, or in the form of a hydrate, solvate, N-oxide orpharmaceutically acceptable salt, as previously described. Typically,such salts are more soluble in aqueous solutions than the correspondingfree acids and bases, but salts having lower solubility than thecorresponding free acids and bases may also be formed.

Pharmaceutical compositions of the invention may take a form suitablefor virtually any mode of administration, including, for example,topical, ocular, oral, buccal, systemic, nasal, injection, transdermal,rectal, vaginal, etc., or a form suitable for administration byinhalation or insufflation.

For topical administration, the active compound(s) or prodrug(s) may beformulated as solutions, gels, ointments, creams, suspensions, etc. asare well-known in the art.

Systemic formulations include those designed for administration byinjection, e.g., subcutaneous, intravenous, intramuscular, intrathecalor intraperitoneal injection, as well as those designed for transdermal,transmucosal oral or pulmonary administration.

Useful injectable preparations include sterile suspensions, solutions oremulsions of the active compound(s) in aqueous or oily vehicles. Thecompositions may also contain formulating agents, such as suspending,stabilizing and/or dispersing agent. The formulations for injection maybe presented in unit dosage form, e.g., in ampules or in multidosecontainers, and may contain added preservatives.

Alternatively, the injectable formulation may be provided in powder formfor reconstitution with a suitable vehicle, including but not limited tosterile pyrogen free water, buffer, dextrose solution, etc., before use.To this end, the active compound(s) may be dried by any art-knowntechnique, such as lyophilization, and reconstituted prior to use.

For transmucosal administration, penetrants appropriate to the barrierto be permeated are used in the formulation. Such penetrants are knownin the art.

For oral administration, the pharmaceutical compositions may take theform of, for example, lozenges, tablets or capsules prepared byconventional means with pharmaceutically acceptable excipients such asbinding agents (e.g., pregelatinised maize starch, polyvinylpyrrolidoneor hydroxypropyl methylcellulose); fillers (e.g., lactose,microcrystalline cellulose or calcium hydrogen phosphate); lubricants(e.g., magnesium stearate, talc or silica); disintegrants (e.g., potatostarch or sodium starch glycolate); or wetting agents (e.g., sodiumlauryl sulfate, lecithin). The tablets may be coated by methods wellknown in the art with, for example, sugars, films or enteric coatings.

Liquid preparations for oral administration may take the form of, forexample, elixirs, solutions, syrups or suspensions, or they may bepresented as a dry product for constitution with water or other suitablevehicle before use. Such liquid preparations may be prepared byconventional means with pharmaceutically acceptable additives such assuspending agents (e.g., sorbitol syrup, cellulose derivatives orhydrogenated edible fats); emulsifying agents (e.g., lecithin oracacia); non-aqueous vehicles (e.g., almond oil, oily esters, ethylalcohol, Cremophore™ or fractionated vegetable oils); and preservatives(e.g., methyl or propyl-p-hydroxybenzoates or sorbic acid). Thepreparations may also contain buffer salts, preservatives, flavoring,coloring and sweetening agents as appropriate.

Preparations for oral administration may be suitably formulated to givecontrolled release of the active compound or prodrug, as is well known.

For buccal administration, the compositions may take the form of tabletsor lozenges formulated in conventional manner.

For rectal and vaginal routes of administration, the active compound(s)may be formulated as solutions (for retention enemas) suppositories orointments containing conventional suppository bases such as cocoa butteror other glycerides.

For nasal administration or administration by inhalation orinsufflation, the active compound(s) or prodrug(s) can be convenientlydelivered in the form of an aerosol spray from pressurized packs or anebulizer with the use of a suitable propellant, e.g.,dichlorodifluoromethane, trichlorofluoromethane,dichlorotetrafluoroethane, fluorocarbons, carbon dioxide or othersuitable gas. In the case of a pressurized aerosol, the dosage unit maybe determined by providing a valve to deliver a metered amount. Capsulesand cartridges for use in an inhaler or insufflator (for examplecapsules and cartridges comprised of gelatin) may be formulatedcontaining a powder mix of the compound and a suitable powder base suchas lactose or starch.

For ocular administration, the active compound(s) or prodrug(s) may beformulated as a solution, emulsion, suspension, etc. suitable foradministration to the eye. A variety of vehicles suitable foradministering compounds to the eye are known in the art. Specificnon-limiting examples are described in U.S. Pat. No. 6,261,547; U.S.Pat. No. 6,197,934; U.S. Pat. No. 6,056,950; U.S. Pat. No. 5,800,807;U.S. Pat. No. 5,776,445; U.S. Pat. No. 5,698,219; U.S. Pat. No.5,521,222; U.S. Pat. No. 5,403,841; U.S. Pat. No. 5,077,033; U.S. Pat.No. 4,882,150; and U.S. Pat. No. 4,738,851.

For prolonged delivery, the active compound(s) or prodrug(s) can beformulated as a depot preparation for administration by implantation orintramuscular injection. The active ingredient may be formulated withsuitable polymeric or hydrophobic materials (e.g., as an emulsion in anacceptable oil) or ion exchange resins, or as sparingly solublederivatives, e.g., as a sparingly soluble salt. Alternatively,transdermal delivery systems manufactured as an adhesive disc or patchwhich slowly releases the active compound(s) for percutaneous absorptionmay be used. To this end, permeation enhancers may be used to facilitatetransdermal penetration of the active compound(s). Suitable transdermalpatches are described in for example, U.S. Pat. No. 5,407,713; U.S. Pat.No. 5,352,456; U.S. Pat. No. 5,332,213; U.S. Pat. No. 5,336,168; U.S.Pat. No. 5,290,561; U.S. Pat. No. 5,254,346; U.S. Pat. No. 5,164,189;U.S. Pat. No. 5,163,899; U.S. Pat. No. 5,088,977; U.S. Pat. No.5,087,240; U.S. Pat. No. 5,008,110; and U.S. Pat. No. 4,921,475.

Alternatively, other pharmaceutical delivery systems may be employed.Liposomes and emulsions are well-known examples of delivery vehiclesthat may be used to deliver active compound(s) or prodrug(s). Certainorganic solvents such as dimethylsulfoxide (DMSO) may also be employed,although usually at the cost of greater toxicity.

The pharmaceutical compositions may, if desired, be presented in a packor dispenser device which may contain one or more unit dosage formscontaining the active compound(s). The pack may, for example, comprisemetal or plastic foil, such as a blister pack. The pack or dispenserdevice may be accompanied by instructions for administration.

5.8 Effective Dosages

The active compound(s) or prodrug(s) of the invention, or compositionsthereof, will generally be used in an amount effective to achieve theintended result, for example in an amount effective to treat or preventthe particular disease being treated. The compound(s) may beadministered therapeutically to achieve therapeutic benefit. Bytherapeutic benefit is meant eradication or amelioration of theunderlying disorder being treated and/or eradication or amelioration ofone or more of the symptoms associated with the underlying disorder suchthat the patient reports an improvement in feeling or condition,notwithstanding that the patient may still be afflicted with theunderlying disorder. Therapeutic benefit also includes halting orslowing the progression of the disease, regardless of whetherimprovement is realized.

The amount of compound administered will depend upon a variety offactors, including, for example, the particular indication beingtreated, the mode of administration, the severity of the indicationbeing treated and the age and weight of the patient, the bioavailabilityof the particular active compound, etc. Determination of an effectivedosage is well within the capabilities of those skilled in the art.

Effective dosages may be estimated initially from in vitro assays. Forexample, an initial dosage for use in animals may be formulated toachieve a circulating blood or serum concentration of active compoundthat is at or above an IC₅₀ of the particular compound as measured in anin vitro assay, such as the in vitro assays described in the Examplessection. Calculating dosages to achieve such circulating blood or serumconcentrations taking into account the bioavailability of the particularcompound is well within the capabilities of skilled artisans. Forguidance, the reader is referred to Fingl & Woodbury, “GeneralPrinciples,” In: Goodman and Gilman's The Pharmaceutical Basis ofTherapeutics, Chapter 1, pp. 1-46, latest edition, Pagamonon Press, andthe references cited therein.

Initial dosages may also be estimated from in vivo data, such as animalmodels. Animal models useful for testing the efficacy of compounds totreat or prevent the various diseases described above are well-known inthe art. Dosage amounts will typically be in the range of from about0.0001 or 0.001 or 0.01 mg/kg/day to about 100 mg/kg/day, but may behigher or lower, depending upon, among other factors, the activity ofthe compound, its bioavailability, the mode of administration andvarious factors discussed above. Dosage amount and interval may beadjusted individually to provide plasma levels of the compound(s) whichare sufficient to maintain therapeutic or prophylactic effect. Forexample, the compounds may be administered once per week, several timesper week (e.g., every other day), once per day or multiple times perday, depending upon, among other things, the mode of administration, thespecific indication being treated and the judgment of the prescribingphysician. In cases of local administration or selective uptake, such aslocal topical administration, the effective local concentration ofactive compound(s) may not be related to plasma concentration. Skilledartisans will be able to optimize effective local dosages without undueexperimentation.

Preferably, the compound(s) will provide therapeutic or prophylacticbenefit without causing substantial toxicity. Toxicity of thecompound(s) may be determined using standard pharmaceutical procedures.The dose ratio between toxic and therapeutic (or prophylactic) LD₅₀/ED₅₀effect is the therapeutic index (LD₅₀ is the dose lethal to 50% of thepopulation and ED₅₀ is the dose therapeutically effective in 50% of thepopulation). Compounds(s) that exhibit high therapeutic indices arepreferred.

5.9 Kits

The compounds and/or prodrugs described herein may be assembled in theform of kits. In some embodiments, the kit provides the compound(s) andreagents to prepare a composition for administration. The compositionmay be in a dry or lyophilized form, or in a solution, particularly asterile solution. When the composition is in a dry form, the reagent maycomprise a pharmaceutically acceptable diluent for preparing a liquidformulation. The kit may contain a device for administration or fordispensing the compositions, including, but not limited to syringe,pipette, transdermal patch, or inhalant.

The kits may include other therapeutic compounds for use in conjunctionwith the compounds described herein. In some embodiments, thetherapeutic agents are other anti-cancer and anti-neoplastic compounds.These compounds may be provided in a separate form, or mixed with thecompounds of the present invention.

The kits will include appropriate instructions for preparation andadministration of the composition, side effects of the compositions, andany other relevant information. The instructions may be in any suitableformat, including, but not limited to, printed matter, videotape,computer readable disk, or optical disc.

6. EXAMPLES 6.1 In Vitro Cellular Experiments

The compounds illustrated in TABLES 1-14, supra, were synthesized usingthe methods described herein. Salts of the compounds were prepared usingstandard techniques.

The IC₅₀ values of various compounds against a various different tumorcell lines were determined using standard in vitro antiproliferationassays. The tumor cell lines tested were as follows: A549 (lung); H1299(lung), ACHN (kidney/p53 wt), CAKI (renal cell carcinoma), NCI-H460(lung); HCT116 (colon/p53wt); HELA (cervix/p53 wt), HUH7 (liver/p53 wt),HUVEC (primary endothelial), LNCAP (prostate), HT-29 (colon); MCF-7(breast); MCF-7 (breast/ER positive), MDA MB435S (breast); MDA MB231(breast/ER negative), DU145 (prostate); BxPC-3 (pancreatic); SKOV-3(ovarian); HepG2 (hepatic), NDHF primary normal human dermalfibroblast), SKMEL28 (breast/p53mut), SKMEL5 (breast/p53 wt), U2OS(bone) and PC3 (prostate). The activity of the compounds against A549cells and H1299 cells is reported in TABLES 1-14, supra. In the tables,a “+” indicates an IC₅₀ of ≦20 μM or less, a “−” indicates an IC₅₀of >20 μM. A value of “−/+” indicates that the specific compoundexhibited an IC₅₀ of >20 μM in a 3-point assay, but exhibited an IC₅₀ of≦20 μM in a higher-point (e.g., 6-point) assay. A value of “+/−”indicates that the specific compound exhibited an IC₅₀ of ≦20 μM in a3-point assay, but exhibited an IC₅₀ of >20 μM in a higher-point (e.g.,6-point) assay. A blank indicates the specified compound was not testedagainst the specified cell line. Many of the compounds tested exhibitedIC50s against A549 and/or H1299 cells in the nanomolar range.

Many of the compounds were tested against other cell lines. Theiractivity is reported in TABLE 15, below, where +”, “−”, “−/+” or “+/−”values are as described for TABLES 1-14. Where the cell line includes avalue in a parenthetical, the value indicates the seeding density atwhich the assay was performed

TABLE 15 No ACHN CAKI HCT116 HELA HUH7 HUVEC LNCAP MCF7(2K) MCF7(4K)MDA-MB-231(2K) MDA-MB-231(4K) NDHF PC3 SKMEL28 SKMEL5 U20S 254 + 256 +300 + 697 + 699 − 710 + 717 − 493 −/+ 494 + 534 + 626 + 495 + 629 +632 + 633 + 634 − 496 − 497 − 732 + 500 + − 636 − 742 + 501 − 502 − 746− 747 + 642 + 748 − 643 − 749 − 750 + 751 − 752 − 503 + − 753 − 754 −755 + 504 + 506 − 507 − 508 − 509 − 644 + − 645 + − 510 − 761 − 762 +763 − 764 − 765 + 511 + 512 + − + + + + + − + + + 766 − 767 − 646 + +− + + + + + + + + 647 + 649 − + + + + + + 516 + + 523 +− + + + + + + + + 790 + 651 − 652 − 305 + 306 − 307 + 835 + + + 309 +310 + 311 + 312 + 313 −/+ 314 −/+ 838 − 527 + 528 + 839 + 841 + 529 +845 + 846 + + + + + + 847 + 848 + 654 + + + + 851 + + +858 + + + + + + + + + + 868 + + + + + + 869 + + + 933 + 938 − 940 + +580 − 581 + 942 + + + + + + + + + + + 947 + 948 + 949 + + + 950 + + 954− 955 − 956 − 957 + 958 − 959 − 960 − 961 − 962 − 355 − + + 356 − 676 +677 − 358 −/+ 359 +/− + 978 − 979 − 980 − 981 + + 982 − 983 − 984 + +985 − 986 − 987 + 418 + 101 + + + + + 102 + + + + + 103 + + + + + 991 +104 − 105 + 106 + + + + + 107 + + − + + + + + + + + + + + + 419 + 420 −110 + 111 + 112 + 113 + 114 + 432 + 435 + 436 + 211 + + + + + +125 + + + + + + 141 + + + + + + 1006 + 1007 − 1009 +

6.2 In Vivo Xenograft Experiments

Compounds 107, 858, 164, 211 and 156 were tested for their ability toshrink tumors generated from A549 cells in standard xenograftexperiments. Compounds 141 and 211 were tested against H1299 tumors.When palpable tumors appeared and were of a preselected volume(approximately 40-150 mm³), the mice were administered test compounds asspecified in TABLE 16, below. Preliminary results are reported in TABLE16.

TABLE 16 Test Preliminary Compound Cell Type Dose Dose ScheduleFormulation Comments Results 107 A549 40 mg/kg 3x a day for 4 LiposomeNo No significant days followed by Vehicle (DMPC) precipitationreduction in 5 days rest. was noted tumor size 107 A549 25 mg/kg andBid, daily 16% Cremophor/ Compound No significant 50 mg/kg And 16% EtOH/crashed out of reduction in Bid 4 days w/8 68% Saline solution whentumor size seen days rest saline was with either added dose 858 A549 25mg/kg Bid, daily 5% EtOH/ No No significant 15% Cremophor/ precipitationreduction in 80% Saline was noted tumor size seen 164, 211 A549 25 mg/kgR563-5 days bid 100% DMSO No 23.2% with 2 days of precipitationreduction in rest was noted tumor size in R565-bid, daily mice treatedwith 211. Compound 164 showed no significant reduction in tumor size.141, 211 H1299 25 mg/kg Bid, daily 100% DMSO Compound 141 No significantprecipitated out reduction in on day 2. tumor size was Compound was seenwith prepared fresh either for days 3, 4, compound. and 5. Noprecipitation was noted for Compound 211 156 A549 25 mg/kg and Bid,daily 0.9% Saline No 60.4% 50 mg/kg precipitation reduction in was notedtumor size was observed in mice treated with 50 mg/kg. No significantreduction was seen at the lower dose.

Although the foregoing inventions have been described in some detail tofacilitate understanding, it will be apparent that certain changes andmodifications may be practiced within the scope of the appended claims.Accordingly, the described embodiments are to be considered asillustrative and not restrictive, and the invention is not to be limitedto the details given herein, but may be modified within the scope andequivalents of the appended claims.

It is to be understood that when ranges of integers are describedherein, the description is intended to include the endpoints and eachintervening integer as though each integer were explicitly delineated.As a specific example, the description “an integer from 1 to 6,” isintended to explicitly describe 1, 2, 3, 4, 5 and 6.

All literature and patent references cited throughout the applicationare incorporated by reference into the application for all purposes.

What is claimed is:
 1. A method of inhibiting proliferation of a cancercell, comprising contacting the cancer cell with an effective amount ofa 2,4-pyrimidinediamine compound according to structural formula (I):

or a prodrug, salt, hydrate, solvate or N-oxides thereof, wherein: L¹and L² are each, independently of one another, selected from a loweralkyldiyl linker, a lower alkylene linker and a covalent bond; R² isselected from the group consisting of lower alkyl optionally substitutedwith an R^(b) group,

 where Y is NH, O or CH₂; R^(2′) is hydrogen, methyl or lower alkyl;R^(4′) is hydrogen, methyl or lower alkyl; R⁴ is

R⁵ is selected from the group consisting of halo, fluoro and —CF₃; R⁶ ishydrogen; each R⁸ is independently selected from the group consisting ofhydrogen, lower alkyl, —(CH₂)_(n)—OH, —OR^(a), —(CH₂)_(n)—NR^(c)R^(c),—O(CH₂)_(n)—R^(a), —O(CH₂)_(n)—R^(b), —C(O)OR^(a), —C(S)O R^(a,) halo,—CF₃ and —OCF₃; each R⁹ is independently selected from the groupconsisting of hydrogen, lower alkyl, —OR^(a), —(CH₂)_(n)—NR^(c)R^(c),—O(CH₂)_(n)—R^(a), —O(CH₂)_(n)—R^(b), —C(O)—NR^(c)R^(c),—C(S)—NR^(c)R^(c), —S(O)₂—NR^(c)R^(c), —NHC(O)R^(a), —NHC(S)R^(a),—C(O)—NH—(CH₂)_(n)—NR^(c)R^(c), —C(S)—NH—(CH₂)_(n)—NR^(c)R^(c), halo,—CF₃, —OCF₃,

each R¹⁰ is independently selected from the group consisting ofhydrogen, lower alkyl, —(CH₂)_(n)—OH, —(CH₂)_(n)—NR^(c)R^(c), —OR^(a),—O(CH₂)_(n)—R^(a), —O(CH₂)_(n)—R^(b), halo, —CF₃, —OCF₃,

each R¹¹ is independently selected from the group consisting of —OR^(a),—NR^(c)R^(c) and —NR^(a)R^(d); each R¹² is independently selected fromthe group consisting of lower alkyl, arylalkyl, —OR^(a), —NR^(c)R^(c),—C(O)R^(a), —C(O)OR^(a) and —C(O)NR^(c)R^(c); each R¹³ is independentlyselected from the group consisting of lower alkyl, hydroxy, loweralkoxy, methoxy, —C(O)NR^(c)R^(c) and —C(O)NH₂; each R¹⁵ isindependently selected from the group consisting of hydrogen, loweralkyl, lower cycloakyl and phenyl; each R¹⁶ is independently selectedfrom the group consisting of hydrogen, methyl, lower alkyl, lowercycloalkyl, lower branched alkyl and lower cycloalkylmethyl; each m isindependently an integer from 1 to 3; each n is independently an integerfrom 1 to 3; each R^(a) is independently selected from the groupconsisting of hydrogen, lower alkyl, lower cycloalkyl, lowercycloalkylalkyl, phenyl and benzyl; each R^(b) is independently selectedfrom the group consisting of —OR^(a), —CF₃, —OCF₃, —NR^(c)R^(c),—C(O)R^(a), —C(S)R^(a), —C(O)OR^(a), —C(S)OR^(a), —C(O)NR^(c)R^(c),—C(S)NR^(c)R^(c), —S(O)₂NR^(c)R^(c), —C(O)NR^(a)R^(d), —C(S)NR^(a)R^(d)and —S(O)₂NR^(a)R^(d); each R^(c) is independently selected from thegroup consisting of hydrogen, lower alkyl and lower cycloalkyl, or,alternatively, two R^(c)s may be taken together with the nitrogen atomto which they are bonded to form a 5-7 membered saturated ring whichoptionally includes 1-2 additional heteroatomic groups selected from O,NR^(a), NR^(a)—C(O)R^(a), NR^(a)—C(O)OR^(a) and NR^(a)—C(O)NR^(a); andeach R^(d) is independently selected from lower mono-hydroxyalkyl andlower di-hydroxyalkyl, with the provisos that: (i) when R² is

 then R⁹ and R¹⁰ are not both simultaneously lower alkoxy or methoxy;(ii) R² is not 3,4,5-trimethoxyphenyl or 3,4,5-tri(loweralkoxy)phenyl;(iii) R² is not lower alkyl; (iv) when R⁴ is

 where R¹⁵ is t-butyl and R² is

 then at least two of R⁸, R⁹ and R¹⁰ or R⁸, R⁹ and R¹³ are other thanhydrogen; and (v) when R⁴ is

 where R¹⁵ is t-butyl and R² is

 where R⁸ and R⁹ are each hydrogen, then R¹⁰ is other than —(CH₂)_(n)—OHor —O(CH₂)_(n)—R^(b) where R^(b) is selected from —NR^(c)R^(c),—C(O)R^(a), —C(O)NR^(c)R^(c) and —C(O)NR^(a)R^(d).
 2. The method ofclaim 1 in which L¹ and L² are each a covalent bond.
 3. The method ofclaim 2 in which R⁵ is fluoro.
 4. The method of claim 3 in which R^(2′)and R^(4′) are each hydrogen.
 5. The method of claim 3 in which R^(2′)is hydrogen and R^(4′) is methyl.
 6. The method of claim 3 in which R²is


7. The method of claim 3 in which R⁴ is


8. The method of claim 7 in which R^(a) is hydrogen and R¹⁵ is selectedfrom lower branched alkyl and lower cycloalkyl.
 9. The method of claim 7in which R¹⁵ is t-butyl or cyclopropyl.
 10. The method of claim 7 inwhich R² is


11. The method of claim 10 in which R⁸ is hydrogen; R⁹ is selected from

and R¹⁰ is other than


12. The method of claim 10 in which R¹⁰ is selected from methyl,trifluoromethyl and chloro.
 13. The method of claim 10 in which R¹² ofR⁹ is selected from methyl, —C(O)CH₃, —C(O)OCH₃ and —C(O)CH₂CH₃.
 14. Themethod of claim 13 in which R⁹ is selected from hydrogen, methyl,trifluoromethyl and chloro.
 15. The method of claim 13 in which R¹² ofR¹⁰ is selected from methyl, —C(O)CH₃, —C(O)OCH₃ and —C(O)CH₂CH₃. 16.The method of claim 10 in which R⁸ is hydrogen; R⁹ is other than

and R¹⁰ is selected from


17. The method of claim 1 wherein the compound is

or a pharmaceutically acceptable salt of any of the foregoing.